70 results on '"Sabine Lüthje"'
Search Results
2. Hypoxia-Induced Aquaporins and Regulation of Redox Homeostasis by a Trans-Plasma Membrane Electron Transport System in Maize Roots
- Author
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Anne Hofmann, Stefanie Wienkoop, and Sabine Lüthje
- Subjects
antioxidant ,aquaporin ,electron transport system ,hypoxia ,plasma membrane ,plant growth regulators ,Therapeutics. Pharmacology ,RM1-950 - Abstract
In plants, flooding-induced oxygen deficiency causes severe stress, leading to growth reduction and yield loss. It is therefore important to understand the molecular mechanisms for adaptation to hypoxia. Aquaporins at the plasma membrane play a crucial role in water uptake. However, their role during hypoxia and membrane redox changes is still not fully understood. The influence of 24 h hypoxia induction on hydroponically grown maize (Zea mays L.) was investigated using an oil-based setup. Analyses of physiological parameters revealed typical flooding symptoms such as increased ethylene and H2O2 levels, an increased alcohol dehydrogenase activity, and an increased redox activity at the plasma membrane along with decreased oxygen of the medium. Transcriptomic analysis and shotgun proteomics of plasma membranes and soluble fractions were performed to determine alterations in maize roots. RNA-sequencing data confirmed the upregulation of genes involved in anaerobic metabolism, biosynthesis of the phytohormone ethylene, and its receptors. Transcripts of several antioxidative systems and other oxidoreductases were regulated. Mass spectrometry analysis of the plasma membrane proteome revealed alterations in redox systems and an increased abundance of aquaporins. Here, we discuss the importance of plasma membrane aquaporins and redox systems in hypoxia stress response, including the regulation of plant growth and redox homeostasis.
- Published
- 2022
- Full Text
- View/download PDF
3. Hypoxia-Responsive Class III Peroxidases in Maize Roots: Soluble and Membrane-Bound Isoenzymes
- Author
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Anne Hofmann, Stefanie Wienkoop, Sönke Harder, Fabian Bartlog, and Sabine Lüthje
- Subjects
aerenchyma ,cell wall remodeling ,class III peroxidases ,hypoxia ,maize roots ,plasma membrane ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Flooding induces low-oxygen environments (hypoxia or anoxia) that lead to energy disruption and an imbalance of reactive oxygen species (ROS) production and scavenging enzymes in plants. The influence of hypoxia on roots of hydroponically grown maize (Zea mays L.) plants was investigated. Gene expression (RNA Seq and RT-qPCR) and proteome (LC–MS/MS and 2D-PAGE) analyses were used to determine the alterations in soluble and membrane-bound class III peroxidases under hypoxia. Gel-free peroxidase analyses of plasma membrane-bound proteins showed an increased abundance of ZmPrx03, ZmPrx24, ZmPrx81, and ZmPr85 in stressed samples. Furthermore, RT-qPCR analyses of the corresponding peroxidase genes revealed an increased expression. These peroxidases could be separated with 2D-PAGE and identified by mass spectrometry. An increased abundance of ZmPrx03 and ZmPrx85 was determined. Further peroxidases were identified in detergent-insoluble membranes. Co-regulation with a respiratory burst oxidase homolog (Rboh) and key enzymes of the phenylpropanoid pathway indicates a function of the peroxidases in membrane protection, aerenchyma formation, and cell wall remodeling under hypoxia. This hypothesis was supported by the following: (i) an elevated level of hydrogen peroxide and aerenchyma formation; (ii) an increased guaiacol peroxidase activity in membrane fractions of stressed samples, whereas a decrease was observed in soluble fractions; and (iii) alterations in lignified cells, cellulose, and suberin in root cross-sections.
- Published
- 2020
- Full Text
- View/download PDF
4. Alterations in Soluble Class III Peroxidases of Maize Shoots by Flooding Stress
- Author
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Claudia-Nicole Meisrimler, Friedrich Buck, and Sabine Lüthje
- Subjects
flooding ,water logging ,guaiacol peroxidases ,native PAGE ,soluble proteins ,shoot ,Zea mays L. ,Microbiology ,QR1-502 - Abstract
Due to changing climate, flooding (waterlogged soils and submergence) becomes a major problem in agriculture and crop production. In the present study, the effect of waterlogging was investigated on peroxidases of maize (Zea mays L.) leaves. The plants showed typical adaptations to flooding stress, i.e., alterations in chlorophyll a/b ratios and increased basal shoot diameter. Seven peroxidase bands could be detected by first dimension modified SDS-PAGE and 10 bands by first dimension high resolution Clear Native Electrophoresis that altered in dependence on plant development and time of waterlogging. Native isoelectric focusing revealed three acidic to neutral and four alkaline guaiacol peroxidases that could be further separated by high resolution Clear Native Electrophorese in the second dimension. One neutral peroxidase (pI 7.0) appeared to be down-regulated within four hours after flooding, whereas alkaline peroxidases (pI 9.2, 8.0 and 7.8) were up-regulated after 28 or 52 h. Second dimensions revealed molecular masses of 133 kDa and 85 kDa for peroxidases at pI 8.0 and 7.8, respectively. Size exclusion chromatography revealed native molecular masses of 30–58 kDa for peroxidases identified as class III peroxidases and ascorbate peroxidases by mass spectrometry. Possible functions of these peroxidases in flooding stress will be discussed.
- Published
- 2014
- Full Text
- View/download PDF
5. International Plant Proteomics Organization (INPPO) World Congress 2014
- Author
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Joshua L Heazlewood, Jesus V Jorrin Novo, Ganesh Kumar Agrawal, Silvia Mazzuca, and Sabine Lüthje
- Subjects
Mass Spectrometry ,plant proteomics ,2-DE ,World congress ,Plant culture ,SB1-1110 - Published
- 2016
- Full Text
- View/download PDF
6. Membrane-Bound Class III Peroxidases: Unexpected Enzymes with Exciting Functions
- Author
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Sabine Lüthje and Teresa Martinez-Cortes
- Subjects
Arabidopsis thaliana ,Class III peroxidase ,Medicago truncatula ,microdomains ,phylogenetics ,plasma membrane ,protein–protein interaction ,Oryza sativa ,tonoplast ,Zea mays ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Class III peroxidases are heme-containing proteins of the secretory pathway with a high redundance and versatile functions. Many soluble peroxidases have been characterized in great detail, whereas only a few studies exist on membrane-bound isoenzymes. Membrane localization of class III peroxidases has been demonstrated for tonoplast, plasma membrane and detergent resistant membrane fractions of different plant species. In silico analysis revealed transmembrane domains for about half of the class III peroxidases that are encoded by the maize (Zea mays) genome. Similar results have been found for other species like thale-cress (Arabidopsis thaliana), barrel medic (Medicago truncatula) and rice (Oryza sativa). Besides this, soluble peroxidases interact with tonoplast and plasma membranes by protein–protein interaction. The topology, spatiotemporal organization, molecular and biological functions of membrane-bound class III peroxidases are discussed. Besides a function in membrane protection and/or membrane repair, additional functions have been supported by experimental data and phylogenetics.
- Published
- 2018
- Full Text
- View/download PDF
7. Proteomic Profiling of the Microsomal Root Fraction: Discrimination of Pisum sativum L. Cultivars and Identification of Putative Root Growth Markers
- Author
-
Claudia-Nicole Meisrimler, Stefanie Wienkoop, and Sabine Lüthje
- Subjects
Pisum sativum ,microsomes ,cultivar comparison ,root morphology ,Microbiology ,QR1-502 - Abstract
Legumes are a large and economically important family, containing a variety of crop plants. Alongside different cereals, some fruits, and tropical roots, a number of leguminosae evolved for millennia as crops with human society. One of these legumes is Pisum sativum L., the common garden pea. In the past, breeding has been largely selective on improved above-ground organs. However, parameters, such as root-growth, which determines acquisition of nutrients and water, have largely been underestimated. Although the genome of P. sativum is still not fully sequenced, multiple proteomic studies have been published on a variety of physiological aspects in the last years. The presented work focused on the connection between root length and the influence of the microsomal root proteome of four different pea cultivars after five days of germination (cultivar Vroege, Girl from the Rhineland, Kelvedon Wonder, and Blauwschokker). In total, 60 proteins were identified to have significantly differential abundances in the four cultivars. Root growth of five-days old seedlings and their microsomal proteome revealed a similar separation pattern, suggesting that cultivar-specific root growth performance is explained by differential membrane and ribosomal protein levels. Hence, we reveal and discuss several putative root growth protein markers possibly playing a key role for improved primary root growth breeding strategies.
- Published
- 2017
- Full Text
- View/download PDF
8. Hypoxia-Responsive Class III Peroxidases in Maize Roots: Soluble and Membrane-Bound Isoenzymes
- Author
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Fabian Bartlog, Sönke Harder, Stefanie Wienkoop, Sabine Lüthje, and Anne Hofmann
- Subjects
0106 biological sciences ,0301 basic medicine ,Proteome ,plasma membrane ,01 natural sciences ,Plant Roots ,lcsh:Chemistry ,Cell Wall ,Gene Expression Regulation, Plant ,Tandem Mass Spectrometry ,Gene expression ,lcsh:QH301-705.5 ,Spectroscopy ,chemistry.chemical_classification ,Phenylpropanoid ,biology ,Chemistry ,food and beverages ,General Medicine ,Cell Hypoxia ,Computer Science Applications ,Isoenzymes ,Biochemistry ,Peroxidases ,class III peroxidases ,aerenchyma ,Oxidation-Reduction ,maize roots ,Peroxidase ,Protein Binding ,respiratory burst oxidase homolog ,Zea mays L ,Zea mays ,Catalysis ,Article ,Aerenchyma ,Aerenchyma formation ,Inorganic Chemistry ,Cell wall ,03 medical and health sciences ,Suberin ,Physical and Theoretical Chemistry ,Molecular Biology ,Reactive oxygen species ,hypoxia ,fungi ,Organic Chemistry ,Cell Membrane ,NADPH Oxidases ,cell wall remodeling ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,biology.protein ,Reactive Oxygen Species ,010606 plant biology & botany ,Chromatography, Liquid - Abstract
Flooding induces low-oxygen environments (hypoxia or anoxia) that lead to energy disruption and an imbalance of reactive oxygen species (ROS) production and scavenging enzymes in plants. The influence of hypoxia on roots of hydroponically grown maize (Zea mays L.) plants was investigated. Gene expression (RNA Seq and RT-qPCR) and proteome (LC&ndash, MS/MS and 2D-PAGE) analyses were used to determine the alterations in soluble and membrane-bound class III peroxidases under hypoxia. Gel-free peroxidase analyses of plasma membrane-bound proteins showed an increased abundance of ZmPrx03, ZmPrx24, ZmPrx81, and ZmPr85 in stressed samples. Furthermore, RT-qPCR analyses of the corresponding peroxidase genes revealed an increased expression. These peroxidases could be separated with 2D-PAGE and identified by mass spectrometry. An increased abundance of ZmPrx03 and ZmPrx85 was determined. Further peroxidases were identified in detergent-insoluble membranes. Co-regulation with a respiratory burst oxidase homolog (Rboh) and key enzymes of the phenylpropanoid pathway indicates a function of the peroxidases in membrane protection, aerenchyma formation, and cell wall remodeling under hypoxia. This hypothesis was supported by the following: (i) an elevated level of hydrogen peroxide and aerenchyma formation, (ii) an increased guaiacol peroxidase activity in membrane fractions of stressed samples, whereas a decrease was observed in soluble fractions, and (iii) alterations in lignified cells, cellulose, and suberin in root cross-sections.
- Published
- 2020
9. In Silico Analysis of Class III Peroxidases: Hypothetical Structure, Ligand Binding Sites, Posttranslational Modifications, and Interaction with Substrates
- Author
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Sabine, Lüthje and Kalaivani, Ramanathan
- Subjects
Isoenzymes ,Binding Sites ,Peroxidases ,Arabidopsis ,Computer Simulation ,Amino Acid Sequence ,Ligands ,Protein Processing, Post-Translational ,Plant Proteins ,Substrate Specificity - Abstract
Functional analyses of peroxidases are a major challenge. In silico analysis appears to be a powerful tool to overcome at least some of the problems that arose from (1) the numerous possible functions of peroxidases, (2) their low substrate specificity, and (3) the compensation of knockout mutants by other isoenzymes. Amino acid sequences and crystal structures of peroxidases were used for the prediction of tertiary structures, posttranslational modifications, ligand and substrate binding sites, and so on of uncharacterized peroxidases. This protocol presents tools and their applications for an in silico analysis of soluble and membrane-bound peroxidases, but it may be used for other proteins, too.
- Published
- 2020
10. In Silico Analysis of Class III Peroxidases: Hypothetical Structure, Ligand Binding Sites, Posttranslational Modifications, and Interaction with Substrates
- Author
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Sabine Lüthje and Kalaivani Ramanathan
- Subjects
0106 biological sciences ,chemistry.chemical_classification ,0303 health sciences ,biology ,Chemistry ,In silico ,Mutant ,Ligand (biochemistry) ,01 natural sciences ,Isozyme ,Protein tertiary structure ,Amino acid ,03 medical and health sciences ,Biochemistry ,biology.protein ,Binding site ,030304 developmental biology ,010606 plant biology & botany ,Peroxidase - Abstract
Functional analyses of peroxidases are a major challenge. In silico analysis appears to be a powerful tool to overcome at least some of the problems that arose from (1) the numerous possible functions of peroxidases, (2) their low substrate specificity, and (3) the compensation of knockout mutants by other isoenzymes. Amino acid sequences and crystal structures of peroxidases were used for the prediction of tertiary structures, posttranslational modifications, ligand and substrate binding sites, and so on of uncharacterized peroxidases. This protocol presents tools and their applications for an in silico analysis of soluble and membrane-bound peroxidases, but it may be used for other proteins, too.
- Published
- 2020
11. Long-term iron deficiency: Tracing changes in the proteome of different pea (Pisum sativum L.) cultivars
- Author
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Sabine Lüthje, Stefanie Wienkoop, Claudia-Nicole Meisrimler, David Lyon, and Christoph-Martin Geilfus
- Subjects
Crops, Agricultural ,0106 biological sciences ,0301 basic medicine ,Proteome ,Iron ,Biophysics ,Plant Roots ,01 natural sciences ,Biochemistry ,Pisum ,03 medical and health sciences ,Sativum ,NDUFA6 ,Gene Expression Regulation, Plant ,Stress, Physiological ,Botany ,Longitudinal Studies ,Cultivar ,Iron deficiency (plant disorder) ,Legume ,Plant Proteins ,biology ,Peas ,food and beverages ,Iron Deficiencies ,Fabaceae ,biology.organism_classification ,Adaptation, Physiological ,030104 developmental biology ,Seeds ,Reactive Oxygen Species ,010606 plant biology & botany - Abstract
Iron deficiency (− Fe) is one of the major problems in crop production. Dicots, like pea ( Pisum sativum L.), are Strategy I plants, which induce a group of specific enzymes such as Fe(III)-chelate reductase (FRO), Fe responsive transporter (IRT) and H + -ATPase (HA) at the root plasma membrane under − Fe. Different species and cultivars have been shown to react diversely to − Fe. Furthermore, different kinds of experimental set-ups for − Fe have to be distinguished: i) short-term vs. long-term, ii) constant vs. acute alteration and iii) buffered vs. unbuffered systems. The presented work compares the effects of constant long-term − Fe in an unbuffered system on roots of four different pea cultivars in a timely manner (12, 19 and 25 days). To differentiate the effects of − Fe and plant development, control plants (+ Fe) were analyzed in comparison to − Fe plants. Besides physiological measurements, an integrative study was conducted using a comprehensive proteome analysis. Proteins, related to stress adaptation (e.g. HSP), reactive oxygen species related proteins and proteins of the mitochondrial electron transport were identified to be changed in their abundance. Regulations and possible functions of identified proteins are discussed. Significance Pea ( Pisum sativum L.) belongs to the legume family (Fabaceae) and is an important crop plant due to high Fe, starch and protein contents. According to FAOSTAT data (September 2015), world production of the garden pea quadrupled from 1970 to 2012. Since the initial studies by Gregor Mendel, the garden pea became the most-characterized legume and has been used in numerous investigations in plant biochemistry and physiology, but is not well represented in the “omics”-related fields. A major limitation in pea production is the Fe availability from soils. Adaption mechanisms to Fe deficiency vary between species, and even cultivars have been shown to react diversely. A label-free proteomic approach, in combination with physiological measurements, was chosen to observe four different pea cultivars for 5 to 25 days. Physiological and proteome data showed that cultivar Blauwschokker and Vroege were more susceptible to − Fe than cultivar Kelvedon (highly efficient) and GftR (semi-efficient). Proteomic data hint that the adaptation process to long-term − Fe takes place between days 19 and 25. Results show that adaptation processes of efficient cultivars are able to postpone secondary negative effects of long-term − Fe, possibly by stabilizing the protein metabolic processing and the mitochondrial electron transport components. This maintains the cellular energy proliferation, keeps ROS production low and postpones the mitochondrial cell death signal.
- Published
- 2016
12. Proteomics of Micronutrient Deficiency and Toxicity
- Author
-
David Hopff, Claudia-Nicole Meisrimler, and Sabine Lüthje
- Subjects
chemistry.chemical_classification ,Reactive oxygen species ,Micronutrient deficiency ,Biochemistry ,Chemistry ,Organelle ,Toxicity ,Transporter ,Micronutrient ,Proteomics ,Redox - Abstract
Micronutrients, such as iron, copper, and zinc are essential for a variety of functions in the living cell. They are essential in different redox systems such as the electron transport chains of cells, organelles, and membranes, but also play an important role in DNA- and RNA-protein interaction. Deprivation of these elements is causing symptoms of deficiency, whereas excess can be toxic due to the production of reactive oxygen species and an imbalance of the cellular redox state. Both deficiency and toxicity are the cause of reduced growth and crop yields. Proteomic approaches revealed alterations of several proteins of different functional classes due to adaptation to nutrient deficiency or toxicity. General and specific responses to micronutrient deficiency and toxicity will be discussed with respect to transporters, signaling, and adaptation mechanisms.
- Published
- 2018
13. Contributors
- Author
-
Surekha Agarwal, Chamizo-Ampudia Alejandro, Llamas Angel, Ana G.L. Assunção, Galvan Aurora, Khurram Bashir, Begoña Blasco, Juan J. Camacho-Cristóbal, Pedro Humberto Castro, Theocharis Chatzistathis, Sardar Alam Cheema, André R. dos Reis, Fernandez Emilio, Muhammad Farooq, Agustín González-Fontes, Kathleen L. Hefferon, María B. Herrera-Rodríguez, David Hopff, Aysha Kiran, Grmay H. Lilay, Sabine Lüthje, Karolina Malas, Satendra K. Mangrauthia, Tejada-Jimenez Manuel, Juan D. Marques Fong, Tsugiyuki Masunaga, Claudia N. Meisrimler, Paloma K. Menguer, Magdalena Migocka, Larissa A.C. Moraes, Adônis Moreira, Niluka Nakandalage, María T. Navarro-Gochicoa, Eloy Navarro-León, Miroslav Nikolic, Levent Ozturk, Jelena Pavlovic, Hafeez ur Rehman, Jesús Rexach, Felipe K. Ricachenevsky, Juan M. Ruiz, Neelamraju Sarla, Saman Seneweera, Raul A. Sperotto, and Abdul Wakeel
- Published
- 2018
14. Isolation and Purity Assessment of Membranes from Norway Spruce
- Author
-
Enni, Väisänen, Junko, Takahashi, Luis A, Jiménez Barboza, Xianbao, Deng, Teemu H, Teeri, Kurt V, Fagerstedt, Sabine, Lüthje, and Anna, Kärkönen
- Subjects
Xylem ,Cell Membrane ,Cell Culture Techniques ,Phloem ,Picea ,Cell Fractionation ,Lignin - Abstract
Gaining membrane vesicles from different plant species and tissue types is crucial for membrane studies. Membrane vesicles can be used for further purification of individual membrane types, and, for example, in studies of membrane enzyme activities, transport assays, and in proteomic analysis. Membrane isolation from some species, such as conifers, has proved to be more difficult than that of angiosperm species. In this paper, we describe steps for isolating cellular membranes from developing xylem, phloem, and lignin-forming tissue-cultured cells of Norway spruce, followed by partial enrichment of plasma membranes by aqueous polymer two-phase partitioning and purity analyses. The methods used are partially similar to the ones used for mono- and dicotyledonous plants, but some steps require discreet optimization, probably due to a high content of phenolic compounds present in the tissues and cultured cells of Norway spruce.
- Published
- 2017
15. Isolation and Purity Assessment of Membranes from Norway Spruce
- Author
-
Xianbao Deng, Sabine Lüthje, Anna Kärkönen, Enni Väisänen, Teemu H. Teeri, Junko Takahashi, Kurt V. Fagerstedt, and Luis Alexis Jiménez Barboza
- Subjects
0106 biological sciences ,0301 basic medicine ,chemistry.chemical_classification ,Aqueous solution ,biology ,Chemistry ,fungi ,food and beverages ,Xylem ,Picea abies ,Isolation (microbiology) ,biology.organism_classification ,01 natural sciences ,03 medical and health sciences ,Tissue culture ,030104 developmental biology ,Enzyme ,Membrane ,Biochemistry ,Botany ,Phloem ,010606 plant biology & botany - Abstract
Gaining membrane vesicles from different plant species and tissue types is crucial for membrane studies. Membrane vesicles can be used for further purification of individual membrane types, and, for example, in studies of membrane enzyme activities, transport assays, and in proteomic analysis. Membrane isolation from some species, such as conifers, has proved to be more difficult than that of angiosperm species. In this paper, we describe steps for isolating cellular membranes from developing xylem, phloem, and lignin-forming tissue-cultured cells of Norway spruce, followed by partial enrichment of plasma membranes by aqueous polymer two-phase partitioning and purity analyses. The methods used are partially similar to the ones used for mono- and dicotyledonous plants, but some steps require discreet optimization, probably due to a high content of phenolic compounds present in the tissues and cultured cells of Norway spruce.
- Published
- 2017
16. Proteomic Profiling of the Microsomal Root Fraction: Discrimination of Pisum sativum L. Cultivars and Identification of Putative Root Growth Markers
- Author
-
Sabine Lüthje, Claudia-Nicole Meisrimler, and Stefanie Wienkoop
- Subjects
0106 biological sciences ,0301 basic medicine ,Clinical Biochemistry ,cultivar comparison ,lcsh:QR1-502 ,01 natural sciences ,Biochemistry ,Article ,lcsh:Microbiology ,Pisum ,Crop ,03 medical and health sciences ,Sativum ,Structural Biology ,Ribosomal protein ,Botany ,Cultivar ,microsomes ,Pisum sativum ,root morphology ,Molecular Biology ,biology ,Proteomic Profiling ,food and beverages ,biology.organism_classification ,030104 developmental biology ,Germination ,Proteome ,010606 plant biology & botany - Abstract
Legumes are a large and economically important family, containing a variety of crop plants. Alongside different cereals, some fruits, and tropical roots, a number of leguminosae evolved for millennia as crops with human society. One of these legumes is Pisum sativum L., the common garden pea. In the past, breeding has been largely selective on improved above-ground organs. However, parameters, such as root-growth, which determines acquisition of nutrients and water, have largely been underestimated. Although the genome of P. sativum is still not fully sequenced, multiple proteomic studies have been published on a variety of physiological aspects in the last years. The presented work focused on the connection between root length and the influence of the microsomal root proteome of four different pea cultivars after five days of germination (cultivar Vroege, Girl from the Rhineland, Kelvedon Wonder, and Blauwschokker). In total, 60 proteins were identified to have significantly differential abundances in the four cultivars. Root growth of five-days old seedlings and their microsomal proteome revealed a similar separation pattern, suggesting that cultivar-specific root growth performance is explained by differential membrane and ribosomal protein levels. Hence, we reveal and discuss several putative root growth protein markers possibly playing a key role for improved primary root growth breeding strategies.
- Published
- 2017
17. International Plant Proteomics Organization (INPPO) World Congress 2014
- Author
-
Jesús V. Jorrín-Novo, Joshua L. Heazlewood, Sabine Lüthje, Silvia Mazzuca, and Ganesh Kumar Agrawal
- Subjects
Engineering ,Workflow ,business.industry ,Event (computing) ,Research community ,Proteome ,Instrumentation (computer programming) ,Proteomics ,Plant biology ,Bioinformatics ,business ,Data science - Abstract
The field of proteomics has advanced considerably over the past two decades. The ability to delve deeper into an organism’s proteome, identify an array of post-translational modifications and profile differentially abundant proteins has greatly expanded the utilization of proteomics. Improvements to instrumentation in conjunction with the development of these reproducible workflows have driven the adoption and application of this technology by a wider research community. However, the full potential of proteomics is far from being fully exploited in plant biology and its translational application needs to be further developed. In 2011, a group of plant proteomic researchers established the International Plant Proteomics Organization (INPPO) to advance the utilization of this technology in plants as well as to create a way for plant proteomics researchers to interact, collaborate and exchange ideas. The INPPO conducted its inaugural world congress in mid 2014 at the University of Hamburg (Germany). Plant proteomic researchers from around the world were in attendance and the event marked the maturation of this research community. The Research Topic captures the opinions, ideas and research discussed at the congress and encapsulates the approaches that were being applied in plant proteomics.The field of proteomics has advanced considerably over the past two decades. The ability to delve deeper into an organism’s proteome, identify an array of post-translational modifications and profile differentially abundant proteins has greatly expanded the utilization of proteomics. Improvements to instrumentation in conjunction with the development of these reproducible workflows have driven the adoption and application of this technology by a wider research community. However, the full potential of proteomics is far from being fully exploited in plant biology and its translational application needs to be further developed. In 2011, a group of plant proteomic researchers established the International Plant Proteomics Organization (INPPO) to advance the utilization of this technology in plants as well as to create a way for plant proteomics researchers to interact, collaborate and exchange ideas. The INPPO conducted its inaugural world congress in mid 2014 at the University of Hamburg (Germany). Plant proteomic researchers from around the world were in attendance and the event marked the maturation of this research community. The Research Topic captures the opinions, ideas and research discussed at the congress and encapsulates the approaches that were being applied in plant proteomics.
- Published
- 2017
18. Alterations in Soluble Class III Peroxidases of Maize Shoots by Flooding Stress
- Author
-
Friedrich Buck, Claudia-Nicole Meisrimler, and Sabine Lüthje
- Subjects
native PAGE ,guaiacol peroxidases ,shoot ,Clinical Biochemistry ,Size-exclusion chromatography ,lcsh:QR1-502 ,Biochemistry ,lcsh:Microbiology ,Article ,Basal shoot ,chemistry.chemical_compound ,flooding ,Structural Biology ,Zea mays L ,Botany ,Molecular Biology ,soluble proteins ,biology ,Chemistry ,Isoelectric focusing ,food and beverages ,Shoot ,biology.protein ,Guaiacol ,water logging ,Ascorbate Peroxidases ,Waterlogging (agriculture) ,Peroxidase - Abstract
Due to changing climate, flooding (waterlogged soils and submergence) becomes a major problem in agriculture and crop production. In the present study, the effect of waterlogging was investigated on peroxidases of maize (Zea mays L.) leaves. The plants showed typical adaptations to flooding stress, i.e., alterations in chlorophyll a/b ratios and increased basal shoot diameter. Seven peroxidase bands could be detected by first dimension modified SDS-PAGE and 10 bands by first dimension high resolution Clear Native Electrophoresis that altered in dependence on plant development and time of waterlogging. Native isoelectric focusing revealed three acidic to neutral and four alkaline guaiacol peroxidases that could be further separated by high resolution Clear Native Electrophorese in the second dimension. One neutral peroxidase (pI 7.0) appeared to be down-regulated within four hours after flooding, whereas alkaline peroxidases (pI 9.2, 8.0 and 7.8) were up-regulated after 28 or 52 h. Second dimensions revealed molecular masses of 133 kDa and 85 kDa for peroxidases at pI 8.0 and 7.8, respectively. Size exclusion chromatography revealed native molecular masses of 30–58 kDa for peroxidases identified as class III peroxidases and ascorbate peroxidases by mass spectrometry. Possible functions of these peroxidases in flooding stress will be discussed.
- Published
- 2014
19. INPPO Actions and Recognition as a Driving Force for Progress in Plant Proteomics: Change of Guard, INPPO Update, and Upcoming Activities
- Author
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Antonio Masi, Abhijit Sarkar, Rungaroon Waditee-Sirisattha, Bongani K. Ndimba, Lello Zolla, Renu Deswal, Jai S. Rohila, Ramesh Sundar Amalraj, Sabine Lüthje, Rainer Cramer, Stefanie Wienkoop, Sixue Chen, Bronwyn J. Barkla, Georgia Tanou, Dominique Job, Yochiro Fukao, Sun Tae Kim, Randeep Rakwal, Tai Wang, Jenny Renaut, Masami Yonekura, Michael J. Dunn, Raj Agrawal, Ganesh Kumar Agrawal, Wolfram Weckwerth, and Thomas Kieselbach
- Subjects
Guard (information security) ,Engineering ,Operations research ,business.industry ,Proteomics ,business ,Molecular Biology ,Biochemistry ,Data science - Abstract
The International Plant Proteomics Organization (INPPO) is a non-profit organization whose members are scientists involved or interested in plant proteomics. Since the publication of the first INPP ...
- Published
- 2013
20. Plasma membrane-associated malate dehydrogenase of maize (Zea mays L.) roots: Native versus recombinant protein
- Author
-
Ljiljana Menckhoff, Friedrich Buck, Mirjana Vuletić, Sabine Lüthje, and Nicole Mielke-Ehret
- Subjects
Proteomics ,Spectrometry, Mass, Electrospray Ionization ,Spinacia ,Molecular Sequence Data ,Biophysics ,Brassica ,medicine.disease_cause ,Plant Roots ,Zea mays ,Biochemistry ,Malate dehydrogenase ,Isozyme ,Malate Dehydrogenase ,Spinacia oleracea ,Tandem Mass Spectrometry ,Protein Interaction Mapping ,Escherichia coli ,medicine ,Amino Acid Sequence ,Disulfides ,Enzyme kinetics ,Glyceraldehyde 3-phosphate dehydrogenase ,Plant Proteins ,Sequence Homology, Amino Acid ,biology ,Gene Expression Profiling ,Cell Membrane ,biology.organism_classification ,Molecular biology ,Recombinant Proteins ,Oxidative Stress ,Ethylmaleimide ,biology.protein ,Spinach ,Heterologous expression ,Protein Processing, Post-Translational - Abstract
Malate dehydrogenase (MDH, EC 1.1.1.37) is involved in several cellular processes including plant development, nutrient uptake and oxidative stress. Evidence for a plasma membrane-associated MDH has been presented for maize (Zea mays L.) roots. In the present study isoenzymes of MDH were purified from highly enriched plasma membrane preparations of maize and compared with soluble isoenzymes (Km, pH optima, pI and molecular masses). Modified SDS-PAGE analyses revealed monomers of 41 kDa for membrane-associated MDH, whereas monomers (35 kDa) and dimers (70 kDa) were detected for soluble isoenzymes. Membrane-associated MDH of cauliflower (Brassica oleracea L.) inflorescences and spinach (Spinacia oleracea L.) leaves showed molecular masses similar to the membrane-associated MDH of maize. The specific maize MDH involved was identified by mass spectrometry (ESI-QTOF-MS/MS, MALDI-TOF-MS). The corresponding gene was cloned and the protein was characterised after heterologous expression in Escherichia coli. Enzyme kinetics and properties of the recombinant and native proteins were compared. The function of thiol groups and the presence of disulphide bonds were analysed by the effect of N-ethylmaleimide, diagonal electrophoresis and labelling. Semiquantitative reverse transcription polymerase chain reaction of maize root transcripts demonstrated a constitutive expression of the gene encoding plasma membrane-associated MDH.
- Published
- 2013
21. Editorial: International Plant Proteomics Organization (INPPO) World Congress 2014
- Author
-
Joshua L. Heazlewood, Silvia Mazzuca, Sabine Lüthje, Ganesh Kumar Agrawal, and Jesús V. Jorrín-Novo
- Subjects
0301 basic medicine ,world congress ,Operations research ,plant proteomics ,Plant Science ,Biology ,2-DE ,Plant biology ,Proteomics ,03 medical and health sciences ,Editorial ,030104 developmental biology ,Research community ,Research questions ,Engineering ethics ,Protein abundance ,mass spectrometry - Abstract
The discipline of proteomics has undergone considerably advances over the past two decades. Our ability to delve deeper into complex proteomes, identify post-translational modifications, and profile protein abundance has greatly expanded the utilization of mass spectrometry in biology. The plant research community has enthusiastically embraced proteomic approaches and has applied these technologies to explore a multitude of research questions in the field of plant biology (Jorrin-Novo et al., 2015). In 2011, a group of plant proteomic researchers established the International Plant Proteomics Organization (INPPO) to advance the application of this technology in plants and agriculture (Agrawal et al., 2011). The INPPO conducted its inaugural world congress in the autumn of 2014 at the University of Hamburg (Germany) (Luthje et al., 2015). The meeting brought together leading international experts in plant proteomics and provided a critical mass for the discussion of proteomic technologies and their application in all aspects of plant biology. This Research Topic arose from this meeting as a means to capture current research, views, and approaches from the wider plant proteomics community.
- Published
- 2016
22. A shotgun proteomic approach reveals that fe deficiency causes marked changes in the protein profiles of plasma membrane and detergent-resistant microdomain preparations from beta vulgaris roots
- Author
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Sabine Lüthje, Ana Flor López-Millán, Elain Gutierrez-Carbonell, Bruno Contreras-Moreira, Sébastien Mongrand, José A. González-Reyes, Matsuo Uemura, Daisuke Takahashi, Anunciación Abadía, Javier Abadía, Estación Experimental de Aula Dei (EEAD) - Plant Stress Physiology Group, Plant Nutrition Department, Spanish National Research Council (CSIC), Fibrostatin SL, University of Valencia, Faculty of Agriculture, United Graduate School of Agricultural Science, Iwate University, Cryobiofrontier Research Center - Faculty of Agriculture, Max Planck Institute of Molecular Plant Physiology (MPI-MP), Max-Planck-Gesellschaft, Biocenter Klein Flottbek, University of Hamburg, Departamento de Biología Celular, Fisiología e Inmunología, University of Córdoba [Córdoba], Biologie végétale intégrative (BVI), Centre National de la Recherche Scientifique (CNRS)-Université Sciences et Technologies - Bordeaux 1-Institut National de la Recherche Agronomique (INRA)-Université Bordeaux Segalen - Bordeaux 2, Laboratoire de biogenèse membranaire (LBM), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Université de Bordeaux (UB), Estación Experimental de Aula Dei (EEAD) Laboratory of Computational and Structural Biology, Consejo Superior de Investigaciones Científicas, Fundación ARAID, Plant Stress Physiology Group - Plant Nutrition Department, Plant Stress Physiology Group, Plant Nutrition Department, Department of Pediatrics, Baylor College of Medicine, USDA-ARS : Agricultural Research Service, Spanish Ministry of Economy and Competitivity (MINECO) [AGL2012-31988, AGL2013-42175-R], FEDER, Aragon Government (Group A03), Japan Society for the Promotion of Science [24-7373, 22120003, 24370018], Bordeaux Metabolome Facility-MetaboHUB [ANR-11-INBS-0010], platform Metabolome-Lipidome-Fluxome of Bordeaux, and JAE Pre-CSIC contract
- Subjects
0106 biological sciences ,0301 basic medicine ,Proteomics ,[SDV]Life Sciences [q-bio] ,Phosphatidic Acids ,Biology ,Plant Roots ,01 natural sciences ,Biochemistry ,Cell membrane ,03 medical and health sciences ,chemistry.chemical_compound ,Membrane Microdomains ,Lipidomics ,medicine ,Phosphorylation ,Iron deficiency (plant disorder) ,Plant Proteins ,Iron deficiency ,Sugar beet ,Cell Membrane ,Lipid microdomain ,Detergent-resistant microdomain ,Iron Deficiencies ,General Chemistry ,Phosphatidic acid ,Lipids ,030104 developmental biology ,Membrane ,medicine.anatomical_structure ,chemistry ,Beta vulgaris ,010606 plant biology & botany ,Plasma membrane - Abstract
15 Pags.- Tabls.- Figs., In the present study we have used label-free shotgun proteomic analysis to examine the effects of Fe deficiency on the protein profiles of highly pure sugar beet root plasma membrane (PM) preparations and detergent-resistant membranes (DRMs), the latter as an approach to study microdomains. Altogether, 545 proteins were detected, with 52 and 68 of them changing significantly with Fe deficiency in PM and DRM, respectively. Functional categorization of these proteins showed that signaling and general and vesicle-related transport accounted for approximately 50% of the differences in both PM and DRM, indicating that from a qualitative point of view changes induced by Fe deficiency are similar in both preparations. Results indicate that Fe deficiency has an impact in phosphorylation processes at the PM level and highlight the involvement of signaling proteins, especially those from the 14–3–3 family. Lipid profiling revealed Fe-deficiency-induced decreases in phosphatidic acid derivatives, which may impair vesicle formation, in agreement with the decreases measured in proteins related to intracellular trafficking and secretion. The modifications induced by Fe deficiency in the relative enrichment of proteins in DRMs revealed the existence of a group of cytoplasmic proteins that appears to be more attached to the PM in conditions of Fe deficiency.
- Published
- 2016
23. Cell wall-bound cationic and anionic class III isoperoxidases of pea root: biochemical characterization and function in root growth
- Author
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Sonja D. Veljovicć-Jovanovicć, Biljana Kukavica, Sabine Lüthje, and Ljiljana Menckhoff
- Subjects
0106 biological sciences ,Anions ,Glycosylation ,Physiology ,IEF–PAGE ,Plant Science ,01 natural sciences ,Plant Roots ,Cell wall ,03 medical and health sciences ,Agglutinin ,Plant Growth Regulators ,Cell Wall ,Cations ,Lectins ,Concanavalin A ,Auxin ,Enzyme Inhibitors ,Polyacrylamide gel electrophoresis ,Pisum sativum ,030304 developmental biology ,Chelating Agents ,Peroxidase ,Plant Proteins ,Gel electrophoresis ,0303 health sciences ,Chitosan ,elicitor ,biology ,Molecular mass ,Indoleacetic Acids ,Isoelectric focusing ,Chemistry ,Peas ,Elicitor ,Isoenzymes ,Molecular Weight ,Kinetics ,Biochemistry ,biology.protein ,pea root ,Electrophoresis, Polyacrylamide Gel ,cell wall peroxidase ,Isoelectric Focusing ,Oxidation-Reduction ,010606 plant biology & botany ,Research Paper - Abstract
Cell wall isolated from pea roots was used to separate and characterize two fractions possessing class III peroxidase activity: (i) ionically bound proteins and (ii) covalently bound proteins. Modified SDS–PAGE separated peroxidase isoforms by their apparent molecular weights: four bands of 56, 46, 44, and 41kDa were found in the ionically bound fraction (iPOD) and one band (70kDa) was resolved after treatment of the cell wall with cellulase and pectinase (cPOD). Isoelectric focusing (IEF) patterns for iPODs and cPODs were significantly different: five iPODs with highly cationic pI (9.5–9.2) were detected, whereas the nine cPODs were anionic with pI values between pH 3.7 and 5. iPODs and cPODs showed rather specific substrate affinity and different sensitivity to inhibitors, heat, and deglycosylation treatments. Peroxidase and oxidase activities and their IEF patterns for both fractions were determined in different zones along the root and in roots of different ages. New iPODs with pI 9.34 and 9.5 were induced with root growth, while the activity of cPODs was more related to the formation of the cell wall in non-elongating tissue. Treatment with auxin that inhibits root growth led to suppression of iPOD and induction of cPOD. A similar effect was obtained with the widely used elicitor, chitosan, which also induced cPODs with pI 5.3 and 5.7, which may be specifically related to pathogen defence. The differences reported here between biochemical properties of cPOD and iPOD and their differential induction during development and under specific treatments implicate that they are involved in specific and different physiological processes. Abbreviations: cPOD covalently bound peroxidase DAB 3,3'-diaminobenzidine DEPMPO spin-trap (5-diethoxy-phosphoryl-5-methyl-1-pyrroline-n-oxide) EPR electron paramagnetic resonance HRP horseradish peroxidase IAA indole-3-acetic acid HRP horseradish peroxidase IEF isoelectric focusing iPOD ionically bound peroxidase NAA naphthalene acetic acid PNGase F peptide N-glycosidase F PR pathogen-related SDS–PAGE sodium dodecyl sulphate–polyacrylamide gel electrophoresis SHAM salicylhydroxamic acid TMB tetramethyl benzidine WGA wheat germ agglutinin
- Published
- 2012
24. Alteration of plasma membrane-bound redox systems of iron deficient pea roots by chitosan
- Author
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Jenny Renaut, Claudia-Nicole Meisrimler, Sébastien Planchon, Kjell Sergeant, and Sabine Lüthje
- Subjects
Vacuolar Proton-Translocating ATPases ,FMN Reductase ,Iron ,Biophysics ,Biology ,Reductase ,Plant Roots ,Biochemistry ,Cell membrane ,Quinone Reductases ,Gene Expression Regulation, Plant ,FMN reductase ,medicine ,Electrophoresis, Gel, Two-Dimensional ,NADH, NADPH Oxidoreductases ,Chitosan ,Gene Expression Profiling ,Cell Membrane ,Peas ,Iron Deficiencies ,Elicitor ,Transport protein ,medicine.anatomical_structure ,Peroxidases ,Proteome ,biology.protein ,Oxidation-Reduction ,Peroxidase - Abstract
Iron is essential for all living organisms and plays a crucial role in pathogenicity. This study presents the first proteome analysis of plasma membranes isolated from pea roots. Protein profiles of four different samples (+Fe, +Fe/Chitosan, -Fe, and -Fe/Chitosan) were compared by native IEF-PAGE combined with in-gel activity stains and DIGE. Using DIGE, 89 proteins of interest were detected in plasma membrane fractions. Data revealed a differential abundance of several spots in all samples investigated. In comparison to the control and -FeCh the abundance of six protein spots increased whereas 56 spots decreased in +FeCh. Altered protein spots were analyzed by MALDI-TOF-TOF mass spectrometry. Besides stress-related proteins, transport proteins and redox enzymes were identified. Activity stains after native PAGE and spectrophotometric measurements demonstrated induction of a ferric-chelate reductase (-Fe) and a putative respiratory burst oxidase homolog (-FeCh). However, the activity of the ferric-chelate reductase decreased in -Fe plants after elicitor treatment. The activity of plasma membrane-bound class III peroxidases increased after elicitor treatment and decreased under iron-deficiency, whereas activity of quinone reductases decreased mostly after elicitor treatment. Possible functions of proteins identified and reasons for a weakened pathogen response of iron-deficient plants were discussed.
- Published
- 2011
25. Diversity of laccases from lichens in suborder Peltigerineae
- Author
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Sabine Lüthje, Richard P. Beckett, Zsanett Laufer, Michael Böttger, and Farida V. Minibayeva
- Subjects
Gene isoform ,Cell wall ,Laccase ,Oxidase test ,Laccase activity ,Tyrosinase ,Botany ,Size-exclusion chromatography ,Plant Science ,Biology ,Lichen ,Ecology, Evolution, Behavior and Systematics - Abstract
Following our previous reports of extracellular laccase activity in lichenized ascomycetes, we investigated the diversity of laccase isoforms in lichens in 20 species from the suborder Peltigerineae. The molecular masses of the active forms of most laccases varied between 135 and 190 kD, although some lichens within the family Peltigeraceae had laccases with higher masses, typically varying from 200 to over 350 kD. Size exclusion chromatography (SEC) was used to confirm the accuracy of the electrophoretic estimates of molecular masses. SEC also clearly distinguished laccase from tyrosinase, another abundant cell wall oxidase in the Peltigerineae. Most species contained one oligomeric laccase isoform. Analysis of replicate collections of four species from different localities showed the isoform a given species contains does not vary with geographical location. The absorption spectra of lichen leachates suggested lichen laccases sometimes resemble the “yellow” laccases found in some free-living fun...
- Published
- 2009
26. Plasma membrane lipid alterations induced by cold acclimation and abscisic acid treatment of winter wheat seedlings differing in frost resistance
- Author
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Ernst Heinz, Petra Sperling, Karl Dörffling, Sabine Lüthje, and Matthias Bohn
- Subjects
biology ,Physiology ,Acclimatization ,Membrane lipids ,fungi ,food and beverages ,Plant Science ,Metabolism ,biology.organism_classification ,Cold Temperature ,Membrane Lipids ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Seedlings ,Seedling ,Cold acclimation ,Osmoprotectant ,Plant hormone ,Agronomy and Crop Science ,Abscisic acid ,Triticum ,Abscisic Acid - Abstract
Cold acclimation of plants affects many aspects of metabolism. Changes in plasma membrane lipids have always been considered to be important for development of frost resistance and survival at subzero temperatures. We studied different cultivars of winter wheat (Triticum aestivum L.) that differed in frost resistance induced either by cold acclimation or treatment with the plant hormone abscisic acid (ABA). Plasma membranes were isolated from non-acclimated and cold- as well as from ABA-acclimated plants, and were subjected to detailed lipid analysis. Cold acclimation affected virtually all plasma membrane lipid components and their constituents, resulting in both increases and decreases, which varied between the three groups of plants investigated. Including the cold-induced variations observed in the few plant species studied in detail previously, cerebrosides were the only components reduced by cold acclimation in all plants. In wheat, more uniform and consistent patterns were obtained when considering colligative parameters such as total free sterols, phospholipids or glycolipids, either as the proportion of total lipids or based on plasma membrane protein. The parameter which changed most significantly in parallel to the increase of inducible frost resistance in the three groups of plants was the ratio of free sterols/glycolipids, which increased. ABA treatment resulted in qualitatively similar effects in only one cultivar, but in general these changes were less pronounced. Compared to changes in transcription rates of several cold-induced genes and in the concentration of various compatible solutes reported for other plants, the observed changes in plasma membrane lipids are minor ones. This may indicate that acclimation-induced changes can be accomplished by posttranscriptional regulation of enzymatic activities, which is in agreement with the failure to detect significant changes in transcription of the corresponding genes during cold induction.
- Published
- 2007
27. Plant-based Foods: Seed, Nutrition and Human Health
- Author
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Sabine Lüthje, Renu Deswal, and Ganesh Kumar Agrawal
- Subjects
Proteomics ,medicine.medical_specialty ,Public health ,MEDLINE ,Plant based ,Biology ,Biochemistry ,Human health ,Environmental health ,Seeds ,medicine ,Humans ,Nutritional Physiological Phenomena ,Food science ,Public Health ,Plants, Edible ,Molecular Biology ,Introductory Journal Article - Published
- 2015
28. Two-dimensional phos-tag zymograms for tracing phosphoproteins by activity in-gel staining
- Author
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Claudia-Nicole Meisrimler, Sabine Lüthje, and Alexandra Schwendke
- Subjects
biology ,Chemistry ,IEF ,High resolution ,Plant Science ,lcsh:Plant culture ,Bioinformatics ,biology.organism_classification ,Enzyme assay ,Staining ,Electrophoresis ,Biochemistry ,NEPHGE ,Phos ,biology.protein ,Posttranslational modification ,Protein phosphorylation ,Phos-tag ,high resolution Clear Native Electrophoresis ,lcsh:SB1-1110 ,in-gel activity staining ,Phosphorylation ,Original Research - Abstract
Protein phosphorylation is one of the most common post translational modifications regulating many cellular processes. The phos-tag technology was combined with two-dimensional zymograms, which consisted of non-reducing IEF PAGE or NEPHGE in the first dimension and high resolution clear native electrophoresis (hrCNE) in the second dimension. The combination of these electrophoresis methods was mild enough to accomplish in-gel activity staining for Fe(III)-reductases by NADH/Fe(III)-citrate/ferrozine, 3,3’-Diaminobenzidine/H2O2 or TMB/H2O2 in the second dimension. The phos-tag zymograms can be used to investigate phosphorylation-dependent changes in enzyme activity. Phos-tag zymograms can be combined with further downstream analysis like mass spectrometry. Non-reducing IEF will resolve proteins with a pI of three to ten, whereas non-reducing NEPHGE finds application for alkaline proteins with a pI higher than eight. Advantages and disadvantages of these new methods will be discussed in detail.
- Published
- 2015
29. INPPO2014, First INPPO World Congress on 'Plant Proteomics: Methodology to Biology'-A global platform for involving, gathering and disseminating knowledge
- Author
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Michael J. Dunn, Abhijit Sarkar, Jenny Renaut, Sebastien Carpentier, Sabine Lüthje, Dominique Job, Raj Agrawal, Randeep Rakwal, Martin Hajduch, Ganesh Kumar Agrawal, Microbiologie, adaptation et pathogénie (MAP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Pole CCMF, Gaz de France Direction Recherche, Gaz De France, Laboratoire Informatique d'Avignon (LIA), Avignon Université (AU)-Centre d'Enseignement et de Recherche en Informatique - CERI, Proteome Research Centre, UCD Conway Institute for Biomolecular and Biomedical Research, University College Dublin [Dublin] (UCD), Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), and Rodrigue, Agnès
- Subjects
Operations research ,[SDV]Life Sciences [q-bio] ,Library science ,[SDV.EE.IEO] Life Sciences [q-bio]/Ecology, environment/Symbiosis ,Cellular level ,Biology ,Plant biology ,Proteomics ,Biochemistry ,[SDV] Life Sciences [q-bio] ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,Community or ,Cost action ,[SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology ,Molecular Biology ,Dissemination ,ComputingMilieux_MISCELLANEOUS ,[SDV.EE.IEO]Life Sciences [q-bio]/Ecology, environment/Symbiosis - Abstract
The International Plant Proteomics Organization (INPPO) is a global platform of the plant proteomics community or, more generally, the scientific community that uses proteomics to address plant biology. Organizing an international conference is one of its initiatives to promote plant proteomics by involving and gathering scientists/researchers/students and by disseminating the acquired knowledge. In this fourth INPPO Highlights, the first INPPO World Congress 2014 (INPPO2014) is described and discussed. The INPPO2014 was held at the University of Hamburg (Germany) with the title "Plant Proteomics: Methodology to Biology" under the leadership of Sabine Luthje (Germany). Participants (around 150) from 38 nations attended this congress covering all continents. The four-day scientific program comprised 52 lectures and 61 poster presentations in a highly professional and friendly atmosphere on mass spectrometry and gel-based proteomics. Two round-table open discussions deliberated on plant proteomics, its associated international organizations/initiatives and future INPPO perspectives. The Second INPPO World Congress 2016 (INPPO2016) "The Quest for Tolerant Varieties-Phenotyping at Plant and Cellular Level" is planned to be organized in Bratislava (Slovakia) under the leadership of Martin Hajduch (Slovak Republic) and Sebastien Carpentier (Belgium) and cosponsored by the COST action FA1306.
- Published
- 2015
30. Reactive oxygen species metabolism in desiccation-stressed thalli of the liverwort Dumortiera hirsuta
- Author
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Richard P. Beckett, Michael Böttger, Farida V. Minibayeva, and Sabine Lüthje
- Subjects
biology ,Physiology ,Superoxide ,Cyanide ,Cell Biology ,Plant Science ,General Medicine ,biology.organism_classification ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Genetics ,Extracellular ,biology.protein ,Desiccation ,Hydrogen peroxide ,Incubation ,Bacteria ,Peroxidase - Abstract
Rates of extracellular superoxide radical formation were estimated in the liverwort Dumortiera hirsuta (SW) Nees. Initial experiments showed that D.hirsuta produced extracellular superoxide at high rates, even when unstressed, and that production increased considerably during rehydration following mild desiccation stress. Experiments in which desiccation was artificially induced using polyethylene glycol showed that superoxide was produced during rehydration rather than desiccation. Incubation of plants in water induced the release about 23% of the superoxide-producing activity, although most appeared to be tightly bound to the cell surface. Experiments with metabolic inhibitors indicated that superoxide production was insensitive to the flavoprotein inhibitor DPI, but inhibited by cyanide, suggesting that peroxidases may produce the superoxide. Despite producing large amounts of superoxide, D. hirsuta only produced small quantities of hydrogen peroxide, the natural product of superoxide dismutation. However, experiments showed that D. hirsuta can reduce the concentration of exogenously supplied hydrogen peroxide from 50μM to zero within 1 h, suggesting that any hydrogen peroxide produced is rapidly metabolized. The physiological significance of superoxide production is discussed, with special reference to its possible role as a defence against pathogenic fungi and bacteria.
- Published
- 2004
31. Transmembrane electron transport in sealed and NAD(P)H-loaded right-side-out plasma membrane vesicles isolated from maize (Zea mays L.) roots
- Author
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Mathias Menckhoff and Sabine Lüthje
- Subjects
Physiology ,chemistry.chemical_element ,Plant Science ,Calcium ,Plant Roots ,Zea mays ,Electron Transport ,chemistry.chemical_compound ,Onium Compounds ,In vivo ,Ferricyanides ,Transport Vesicles ,Edetic Acid ,Vesicle ,Cell Membrane ,Biological Transport ,Electron transport chain ,Transmembrane protein ,Membrane ,chemistry ,Biochemistry ,Biophysics ,Warfarin ,NAD+ kinase ,Ferricyanide ,NADP - Abstract
Electron transport across plasma membranes has been observed in vivo in several plant species and tissues after the application of ferricyanide (hexacyanoferrate III, HCF III). In the present work, a transmembrane electron flow was demonstrated in sealed and NAD(P)H-loaded right-side-out (apoplastic-side-out) plasma membrane vesicles isolated from maize (Zea mays L.) roots. HCF III was reduced at a rate of up to 126 nmol min(-1) mg(-1) protein by NADPH-loaded vesicles, while reduction rates with NADH-loaded vesicles were several-fold lower. Coincident with the reduction of HCF III, NAD(P)H oxidation was observed inside the vesicles. The dependence of reduction on K+ indicated an electrogenic transmembrane electron flow. Application of 100 microM calcium decreased HCF III reduction up to 66%, while pre-incubation with 200 microM warfarin or diphenylene iodonium inhibited transmembrane electron transport only weakly. Fe(3+)-EDTA was not reduced significantly by NADPH-loaded plasma membrane vesicles, whereas XTT was reduced at a rate of 765 pmol min(-1) mg(-1) protein. The results suggested a major function for NADPH in transmembrane electron flow and were discussed in conjunction with in vivo experiments.
- Published
- 2004
32. Possible functions of extracellular peroxidases in stress-induced generation and detoxification of active oxygen species
- Author
-
Angela Mika, Sabine Lüthje, Richard P. Beckett, and Farida V. Minibayeva
- Subjects
biology ,Superoxide ,fungi ,food and beverages ,Plant Science ,Apoplast ,Respiratory burst ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Oxidative enzyme ,Oxidizing agent ,Extracellular ,biology.protein ,Hydrogen peroxide ,Biotechnology ,Peroxidase - Abstract
Extracellular peroxidases are classified as free, or ionically or covalently bound to the cell wall. In addition, peroxidase-like activities have often been demonstrated at the outer surface of protoplasts and plasma membrane preparations. Under certain conditions apoplastic peroxidases have been shown to contribute to the formation of superoxide and hydrogen peroxide during the `oxidative burst' through the oxidation of a reductant. However, the identity of this reductant remains unclear. It has been suggested that the production of these active oxygen species may play important roles in plant responses to biotic and abiotic stress. Extracellular release of pre-existing and de novo synthesis of apoplastic peroxidases is regulated by changing environmental conditions. While the oxidative burst could potentially be harmful to a plant's own cells, tissues can rapidly metabolize even high concentrations of hydrogen peroxide. Recent work has shown that when extracellular hydrogen peroxide exceeds the supplies of reductants, class II and class III peroxidases can display catalase-like activity. Under these conditions, hydrogen peroxide is able to act as both oxidizing and reducing substrate. It seems likely therefore, that a further role of extracellular peroxidases is to protect plants from the consequences of the oxidative burst that they themselves are responsible for producing.
- Published
- 2004
33. Properties of Guaiacol Peroxidase Activities Isolated from Corn Root Plasma Membranes
- Author
-
Angela Mika and Sabine Lüthje
- Subjects
chemistry.chemical_classification ,Gel electrophoresis ,Chromatography ,biology ,Molecular mass ,Physiology ,Size-exclusion chromatography ,Ion chromatography ,Plant Science ,Cofactor ,chemistry.chemical_compound ,Enzyme ,chemistry ,Biochemistry ,Genetics ,biology.protein ,Heme ,Peroxidase - Abstract
Although several investigations have demonstrated a plasma membrane (PM)-bound peroxidase activity in plants, this study is the first, to our knowledge, to purify and characterize the enzymes responsible. Proteins were extracted from highly enriched and thoroughly washed PMs. Washing and solubilization procedures indicated that the enzymes were tightly bound to the membrane. At least two distinct peroxidase activities could be separated by cation exchange chromatography (pmPOX1 and pmPOX2). Prosthetic groups were identified in fractions with peroxidase activity by absorption spectra, and the corresponding protein bands were identified by heme staining. The activities of the peroxidase enzymes responded different to various substrates and effectors and had different thermal stabilities and pH and temperature optima. Because the enzymes were localized at the PM and were not effected by p-chloromercuribenzoate, they were probably class III peroxidases. Additional size exclusion chromatography of pmPOX1 revealed a single activity peak with a molecular mass of 70 kD for the native enzyme, whereas pmPOX2 had two activity peaks (155 and 40 kD). Further analysis of these fractions by a modified sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with heme staining confirmed the estimated molecular masses of the size exclusion chromatography.
- Published
- 2003
34. Salicylic acid changes the properties of extracellular peroxidase activity secreted from wounded wheat ( Triticum aestivum L.) roots
- Author
-
Sabine Lüthje, Farida V. Minibayeva, and Angela Mika
- Subjects
Ion chromatography ,chemistry.chemical_element ,Plant Science ,Calcium ,Plant Roots ,chemistry.chemical_compound ,Extracellular ,Triticum ,Peroxidase ,Plant Diseases ,chemistry.chemical_classification ,biology ,Cell Biology ,General Medicine ,Adaptation, Physiological ,Apoplast ,Enzyme Activation ,Isoenzymes ,Enzyme ,chemistry ,Biochemistry ,biology.protein ,Specific activity ,Extracellular Space ,Salicylic Acid ,Salicylic acid - Abstract
Wheat ( Triticum aestivum L.) roots released proteins showing peroxidase activity in the apoplastic solution in response to wound stress. Preincubation of excised roots with 1 mM salicylic acid at pH 7.0 enhanced the guaiacol peroxidase activity of the extracellular solution (so-called extracellular peroxidase). The soluble enzymes were partially purified by precipitation with ammonium sulfate followed by size exclusion and ion exchange chromatography. Despite an increase in the total activity of secreted peroxidase induced by pretreatment of excised roots with salicylic acid, the specific activity of the partially purified protein was significantly lower compared to that of the control. Purification of the corresponding proteins by ion exchange chromatography indicates that several isoforms of peroxidase occurred in both control and salicylic acid-treated samples. The activities of the extracellular peroxidases secreted by the salicylic acid-treated roots responded differently to calcium and lectins compared with those from untreated roots. Taken together, our data suggest that salicylic acid changes the isoforms of peroxidase secreted by wounded wheat roots.
- Published
- 2003
35. Isolation of cellular membranes from lignin-producing tissues of Norway spruce and analysis of redox enzymes
- Author
-
Teresa Laitinen, Kurt V. Fagerstedt, Anna Kärkönen, Junko Takahashi, Sadette Salonvaara, Sami Holmström, Sabine Lüthje, Luis Alexis Jiménez Barboza, Enni Väisänen, Claudia-Nicole Meisrimler, and Stefanie Wienkoop
- Subjects
Physiology ,Plant Science ,Lignin ,Superoxide dismutase ,chemistry.chemical_compound ,Cytochrome P-450 Enzyme System ,Superoxides ,Xylem ,Cytochrome b5 ,Genetics ,Extracellular ,Picea ,Peroxidase ,Plant Proteins ,chemistry.chemical_classification ,biology ,Superoxide ,Superoxide Dismutase ,Cell Membrane ,Cell Biology ,General Medicine ,Hydrogen Peroxide ,Catalase ,NAD ,Membrane ,Enzyme ,chemistry ,Biochemistry ,Peroxidases ,biology.protein ,Oxidation-Reduction ,Juglone ,NADP - Abstract
There are no earlier reports with successful isolation of plasma membranes from lignin-forming tissues of conifers. A method to isolate cellular membranes from extracellular lignin-producing tissue-cultured cells and developing xylem of Norway spruce was optimized. Modifications to the homogenization buffer were needed to obtain membranes from these phenolics-rich tissues. Membranes were separated by aqueous polymer two-phase partitioning. Chlorophyll a determination, marker enzyme assays and western blot analyses using antibodies for each membrane type showed that mitochondrial, chloroplastic and to a certain extent also ER and Golgi membranes were efficiently diminished from the upper phase, but tonoplast and plasma membranes distributed evenly between the upper and lower phases. Redox enzymes present in the partially purified membrane fractions were assayed in order to reveal the origin of H(2)O(2) needed for lignification. The membranes of spruce contained enzymes able to generate superoxide in the presence of NAD(P)H. Besides members of the flavodoxin and flavodoxin-like family proteins, cytochrome b5, cytochrome P450 and several stress responsive proteins were identified by nitroblue tetrazolium staining of isoelectric focusing gels and by mass spectrometry. Naphthoquinones juglone and menadione increased superoxide production in activity-stained gels. Some juglone-activated enzymes were preferentially using NADH. With NADH, menadione activated only some of the enzymes that juglone did, whereas with NADPH the activation patterns were identical. Duroquinone, a benzoquinone, did not affect superoxide production. Superoxide dismutase, ascorbate peroxidase, catalase and an acidic class III peroxidase isoenzyme were detected in partially purified spruce membranes. The possible locations and functions of these enzymes are discussed.
- Published
- 2014
36. Interaction between electron transport at the plasma membrane and nitrate uptake by maize (Zea mays L.) roots
- Author
-
Olaf Döring, Michael Böttger, M. A. Busch, Sabine Lüthje, and D Steffen
- Subjects
Dicumarol ,Plant Science ,Reductase ,Nitrate reductase ,Nitrate Reductase ,Plant Roots ,Zea mays ,Electron Transport ,chemistry.chemical_compound ,Nitrate ,Nitrate Reductases ,Oxidoreductase ,NADH, NADPH Oxidoreductases ,Nitrite ,Ferricyanides ,Incubation ,Plant Proteins ,chemistry.chemical_classification ,Nitrates ,Uncoupling Agents ,Cell Membrane ,Biological Transport ,Cell Biology ,General Medicine ,Hydrogen-Ion Concentration ,Membrane ,Biochemistry ,chemistry ,Ferricyanide ,Oxidation-Reduction - Abstract
In the present study nitrate uptake by maize (Zea mays L.) roots was investigated in the presence or absence of ferricyanide (hexacyanoferrate III) or dicumarol. Nitrate uptake caused an alkalization of the medium. Nitrate uptake of intact maize seedlings was inhibited by ferricyanide while the effect of dicumarol was not very pronounced. Nitrite was not detected in the incubation medium, neither with dicumarol-treated nor with control plants after application of 100 microM nitrate to the incubation solution. In a second set of experiments interactions between nitrate and ferricyanide were investigated in vivo and in vitro. Nitrate (1 or 3 mM) did neither influence ferricyanide reductase activity of intact maize roots nor NADH-ferricyanide oxidoreductase activity of isolated plasma membranes. Nitrate reductase activity of plasma-membrane-enriched fractions was slightly stimulated by 25 microM dicumarol but was not altered by 100 microM dicumarol, while NADH-ferricyanide oxidoreductase activity was inhibited in the presence of dicumarol. These data suggest that plasma-membrane-bound standard-ferricyanide reductase and nitrate reductase activities of maize roots may be different. A possible regulation of nitrate uptake by plasmalemma redox activity, as proposed by other groups, is discussed.
- Published
- 2001
37. Inhibition of trans-membrane hexacyanoferrate III reductase activity and proton secretion of maize (Zea mays L.) roots by thenoyltrifluoroacetone
- Author
-
Sabine Lüthje and Olaf Döring
- Subjects
Dicumarol ,Plant Science ,Plant Roots ,Zea mays ,Redox ,chemistry.chemical_compound ,Proton transport ,NADH, NADPH Oxidoreductases ,Enzyme Inhibitors ,Ferricyanides ,Thenoyltrifluoroacetone ,Chelating Agents ,Uncoupling Agents ,Cell Membrane ,Cell Biology ,General Medicine ,Hydrogen-Ion Concentration ,Electron transport chain ,Apoplast ,Membrane ,chemistry ,Biochemistry ,Warfarin ,Ferricyanide ,NAD+ kinase ,Protons ,Oxidation-Reduction ,Nuclear chemistry - Abstract
Intact plants can reduce external oxidants by an appearingly trans-membrane electron transport. In vivo an increase in net medium acidification accompanies the reduction of the apoplastic substrate. Up to now, several NAD(P)H dehydrogenases, b-type cytochromes, and a phylloquinone have been identified and partially purified from plant plasma membranes. The occurrence of a quinone in the plasma membrane of maize roots supports the hypothetical model of a proton-transferring redox system, i.e., an electron transport chain with a quinone as mobile electron and proton carrier. In the present study the trans-membrane electron transport system of intact maize (Zea mays L.) roots was investigated. Flow-through and ionostat systems have been used to estimate the electron and proton transport activity of this material. Application of 4,4,4-trifluoro-1-(2-thienyl)-butane-1,3-dione (thenoyltrifluoroacetone) inhibited the reduction of ferricyanide in the incubation solution of intact maize roots up to 70%. This inhibition could not be washed off by rinsing the roots with fresh incubation medium. The acidification of the medium induced after ferricyanide application was inhibited to about 62%. The effects of thenoyltrifluoroacetone on proton fluxes in the absence of ferricyanide have been characterized in a pH-stat system. The net medium acidification by maize roots was inhibited up to 75% by thenoyltrifluoroacetone in the absence of ferricyanide, while dicumarol inhibited net acidification completely. The inhibition of H(+)-ATPase activity was estimated with plasma membrane vesicles isolated by phase partitioning and treated with 0.05% (w/v) Brij 58. ATP-dependent proton gradients and Pi release were measured after preincubation with the effectors. The proton pumping activity by those plasma membrane vesicles was inhibited by dicumarol (53.6%) and thenoyltrifluoroacctone (77.8%), while the release of Pi was unaffected by both inhibitors.
- Published
- 2001
38. Class III peroxidases
- Author
-
Sabine, Lüthje, Claudia-Nicole, Meisrimler, David, Hopff, Tim, Schütze, Jenny, Köppe, and Katrin, Heino
- Subjects
Proteomics ,Peroxidases ,Solubility ,Cell Membrane ,Vacuoles ,Electrophoresis, Gel, Two-Dimensional ,Cell Fractionation ,Zea mays ,Plant Proteins - Abstract
Class III peroxidases are heme-containing proteins of the secretory pathway with an extremely high number of isoenzymes, indicating the tremendous and important functions of this protein family. This chapter describes fractionation of the cell in subproteomes, their separation by polyacrylamide gel electrophoresis (PAGE) and visualization of peroxidase isoenzymes by heme and specific in-gel staining procedures. Soluble and membrane-bound peroxidases were separated by differential centrifugation. Aqueous polymer two-phase partitioning and discontinuous sucrose density gradient were applied to resolve peroxidase profiles of plasma membranes and tonoplast. Peroxidase isoenzymes of subproteomes were further separated by PAGE techniques such as native isoelectric focussing (IEF), high resolution clear native electrophoresis (hrCNE), and modified sodium dodecyl sulfate (modSDS)-PAGE. These techniques were used as stand-alone method or in combination for two-dimensional PAGE.
- Published
- 2013
39. Quinones in plant plasma membranes — a missing link?
- Author
-
J. M. Villalba, Sabine Lüthje, Han Asard, J. A. González-Reyes, P. Van Gestelen, María del Carmen Córdoba-Pedregosa, and Michael Böttger
- Subjects
Flavin adenine dinucleotide ,Chromatography ,biology ,Flavin mononucleotide ,Flavoprotein ,Cell Biology ,Plant Science ,General Medicine ,Flavin group ,Ascorbic acid ,Naphthoquinone ,chemistry.chemical_compound ,Membrane ,NAD(P)H dehydrogenase ,chemistry ,Biochemistry ,biology.protein - Abstract
The occurrence of a vitamin-K-like substance (naphthoquinone group) and flavins (flavin mononucleotide and flavin adenine dinucleotide) is demonstrated in plasma membranes isolated from maize (Zea mays L.) roots, on the basis of high-pressure liquid chromotography and spectral analysis. At least three NAD(P)H dehydrogenases could be purified to homogeneity from this plant material. Two of these proteins (25 and 30 kDa) reduce hexacyanoferrate III and quinones, while the third (41 kDa) reduces oxalacetic acid but not hexacyanoferrate III in the presence of NADH. Low-temperature spectra demonstrate the occurrence of a b-type cytochrome in plasma membranes isolated from maize roots. The latter compound could be reduced by ascorbic acid (E0′ > +80 mV) and shows an α-band maximum at 559 nm (at −196 °C). NADH-dependent cytochromeb reduction could be observed only in the presence of detergent and increased after preincubation with vitamin K3 (menadione). On the basis of the presented data a possible function of naphthoquinones in plasma membrane electron transfer is discussed.
- Published
- 1998
40. Oxidoreductases in plant plasma membranes
- Author
-
Michael Böttger, Hartwig Lüthen, Sabine Lüthje, Sigrid Heuer, and Olaf Döring
- Subjects
chemistry.chemical_classification ,Cell Membrane ,Biophysics ,Biological Transport ,Biological membrane ,Cell Biology ,Hydrogen-Ion Concentration ,Plants ,Biology ,Cytosol ,Membrane ,Iron reduction ,Biochemistry ,chemistry ,Oxidoreductase ,High activity ,Signal transduction ,Oxidoreductases ,Oxidation-Reduction ,Function (biology) ,Ion channel - Abstract
Electron transporting oxidoreductases at biological membranes mediate several physiological processes. While such activities are well known and widely accepted as physiologically significant for other biological membranes, oxidoreductase activities found at the plasma membrane of plants are still being neglected. The ubiquity of the oxidoreductases in the plasma membrane suggests that the activity observed is of major importance in fact up to now no plant without redox activity at the plasmalemma is known. Involvement in proton pumping, membrane energization, ion channel regulation, iron reduction, nutrient uptake, signal transduction, and growth regulation has been proposed. However, positive proof for one of the numerous theories about the physiological function of the system is still missing. Evidence for an involvement in signalling and regulation of growth and transport activities at the plasma membrane is strong, but the high activity of the system displayed in some experiments also suggests function in defense against pathogens.
- Published
- 1997
41. Class III Peroxidases
- Author
-
Sabine Lüthje, Claudia-Nicole Meisrimler, Jenny Köppe, Tim Schütze, Katrin Heino, and David Hopff
- Subjects
Differential centrifugation ,Electrophoresis ,chemistry.chemical_compound ,Isoelectric point ,Density gradient ,Biochemistry ,biology ,Chemistry ,biology.protein ,Sodium dodecyl sulfate ,Polyacrylamide gel electrophoresis ,Heme ,Peroxidase - Abstract
Class III peroxidases are heme-containing proteins of the secretory pathway with an extremely high number of isoenzymes, indicating the tremendous and important functions of this protein family. This chapter describes fractionation of the cell in subproteomes, their separation by polyacrylamide gel electrophoresis (PAGE) and visualization of peroxidase isoenzymes by heme and specific in-gel staining procedures. Soluble and membrane-bound peroxidases were separated by differential centrifugation. Aqueous polymer two-phase partitioning and discontinuous sucrose density gradient were applied to resolve peroxidase profiles of plasma membranes and tonoplast. Peroxidase isoenzymes of subproteomes were further separated by PAGE techniques such as native isoelectric focussing (IEF), high resolution clear native electrophoresis (hrCNE), and modified sodium dodecyl sulfate (modSDS)-PAGE. These techniques were used as stand-alone method or in combination for two-dimensional PAGE.
- Published
- 2013
42. Molecular components and biochemistry of electron transport in plant plasma membranes (Review)
- Author
-
Sabine Lüthje and Olaf Döring
- Subjects
biology ,Chemistry ,Radical ,Cell Membrane ,Cell Biology ,Plasma ,Plants ,Electron transport chain ,Electron Transport ,Chloroplast ,Electron transfer ,Mitochondrial membrane transport protein ,Membrane ,Biochemistry ,biology.protein ,Molecule ,Molecular Biology - Abstract
SummaryIt is worthwhile emphasizing the importance of electron transport across lipid membranes. Mitochondrial and electron transport in chloroplasts were elucidated in great detail many years ago. Plasma membrane-bound electron transfer may be involved in several processes such as membrane energization, signalling, regulation of transport and/or growth, and generation or scavenging of free radicals. We here give an overview of plasma membrane-bound electron transfer, of possible compounds of the electron transporting systems isolated from plasma membranes, and of their biochemical characteristics. Both the progress made in purification of redox enzymes and compounds and data from biochemical characterization of the activities found, support the discussion concerning models of the molecular structure of the electron transport systems of plant plasma membranes.
- Published
- 1996
43. Plasma membrane electron pathways and oxidative stress
- Author
-
Sabine Lüthje, Kerstin Wöltje, Benjamin Möller, and François Clement Perrineau
- Subjects
Cytochrome b561 ,Physiology ,Chemistry ,Clinical Biochemistry ,Cell Membrane ,Electrons ,Cell Biology ,medicine.disease_cause ,Biochemistry ,Redox ,Apoplast ,Cytosol ,Oxidative Stress ,Quinone Reductases ,medicine ,General Earth and Planetary Sciences ,NAD+ kinase ,Signal transduction ,Molecular Biology ,Oxidative stress ,General Environmental Science - Abstract
Several redox compounds, including respiratory burst oxidase homologs (Rboh) and iron chelate reductases have been identified in animal and plant plasma membrane (PM). Studies using molecular biological, biochemical, and proteomic approaches suggest that PM redox systems of plants are involved in signal transduction, nutrient uptake, transport, and cell wall-related processes. Function of PM-bound redox systems in oxidative stress will be discussed.Present knowledge about the properties, structures, and functions of these systems are summarized. Judging from the currently available data, it is likely that electrons are transferred from cytosolic NAD(P)H to the apoplast via quinone reductases, vitamin K, and a cytochrome b561. In tandem with these electrons, protons might be transported to the apoplastic space.Recent studies suggest localization of PM-bound redox systems in microdomains (so-called lipid or membrane rafts), but also organization of these compounds in putative and high molecular mass protein complexes. Although the plant flavocytochrome b family is well characterized with respect to its function, the molecular mechanism of an electron transfer reaction by these compounds has to be verified. Localization of Rboh in other compartments needs elucidation.Plant members of the flavodoxin and flavodoxin-like protein family and the cytochrome b561 protein family have been characterized on the biochemical level, postulated localization, and functions of these redox compounds need verification. Compositions of single microdomains and interaction partners of PM redox systems have to be elucidated. Finally, the hypothesis of an electron transfer chain in the PM needs further proof.
- Published
- 2012
44. The role of active oxygen in iron tolerance of rice (Oryza sauva L.)
- Author
-
K. Bode, Sabine Lüthje, H. U. Neue, M. Böttger, and Olaf Döring
- Subjects
Oryza sativa ,biology ,Chemistry ,Bronzing ,food and beverages ,Hydroculture ,Cell Biology ,Plant Science ,General Medicine ,Oryza ,biology.organism_classification ,Superoxide dismutase ,Horticulture ,Catalase ,Botany ,Shoot ,biology.protein ,Peroxidase - Abstract
Iron tolerance of rice (Oryza sativa L.) was investigated using an oxygen depleted hydroculture system. Treatment with high concentrations of Fe2+ induced yellowing and bronzing symptoms as well as iron coatings at the root surface. Root and shoot growth were inhibited by increasing iron concentration in the medium. All symptoms were more pronounced in an iron sensitive cultivar (IR 64) compared to an iron tolerant one (IR 9764-45-2). Superoxide dismutase and peroxidase activity of root extracts of IR 97 were about twice that of IR 64 in untreated control plants. No significant increase of peroxidase activity was detected with increasing iron concentration in the medium. Catalase activity of IR 64 was slightly higher than that of IR 97, independent of iron concentration.
- Published
- 1995
45. Iron and copper in plasma membranes of maize (Zea mays L.) roots investigated by proton induced X-ray emission
- Author
-
Michael Böttger, M. Niecke, and Sabine Lüthje
- Subjects
Aqueous solution ,Proton ,X-ray ,chemistry.chemical_element ,Cell Biology ,Plant Science ,General Medicine ,Plasma ,Zinc ,Copper ,Membrane ,chemistry ,Elemental analysis ,Botany ,Nuclear chemistry - Abstract
Plasma membranes of maize (Zea mays L., cv. Sil Anjou 18) roots were isolated by aqueous two-phase partitioning. Multi elemental analysis by proton induced X-ray emission (PIXE) was used for the investigation of elemental composition of plasma membranes. Fe, Cu, and Zn as well as P, S, and Ca were identified. We did not find significant amounts of V, Mn, Se, Mo, or W.
- Published
- 1995
46. Changes in the glutathione level induced by transplasma membrane electron transport in maize (Zea mays L.)
- Author
-
Sabine Lüthje, Michael Böttger, S. Krüger, S. Seidenberg, and Olaf Döring
- Subjects
chemistry.chemical_classification ,Cell Biology ,Plant Science ,General Medicine ,Glutathione ,Electron acceptor ,Electron transport chain ,chemistry.chemical_compound ,Electron transfer ,Coleoptile ,chemistry ,Biochemistry ,Biosynthesis ,Thiol ,Cysteine - Abstract
We investigated changes of thiols (GSH, GSSG, and cysteine) induced by transplasma membrane electron transport after addition of artificial electron acceptors and the influence of the thiol level on redox activity. GSH, GSSG, and cysteine content of maize (Zea mays L. cv. Golden Bantam) roots and coleoptile segments was determined by high performance liquid chromatography with a fluorescence detector. GSSG increased after treatment with 0.8 mM diamide, an SH-group oxidizer. GSH level of roots increased after treatment with diamide, while GSH levels of coleoptiles decreased. Incubation of roots with the GSH biosynthesis inhibitor buthionine-D,L-sulfoximine for 6 days lowered the glutathione level up to 80%. However, the GSH/GSSG ratio of maize roots remained constant after treatment with both effectors. The GSH/GSSG ratio and the glutathione level were changed by addition of artificial electron acceptors like hexacyanoferrate (III) or hexabromoiridate (IV), which do not permeate the plasma membrane. Hexacyanoferrate (III) reduction was inhibited up to 25% after the cellular glutathione level was lowered by treatment with diamide or buthionine-D,L-sulfoximine. Proton secretion induced by reduction of the electron acceptors was not affected by both modulators. The change in glutathione level is different for roots and coleoptiles. Our data are discussed with regard to the role of GSH in electron donation for a plasma membrane bound electron transport system.
- Published
- 1995
47. IPG-strips versus off-gel fractionation: advantages and limits of two-dimensional PAGE in separation of microsomal fractions of frequently used plant species and tissues
- Author
-
Claudia-Nicole Meisrimler and Sabine Lüthje
- Subjects
Proteomics ,Biophysics ,Arabidopsis ,Fractionation ,Chemical Fractionation ,Biochemistry ,Plant Roots ,Zea mays ,Pisum ,Sativum ,Microsomes ,Tobacco ,Chemical Precipitation ,Electrophoresis, Gel, Two-Dimensional ,Plant Proteins ,Two-dimensional gel electrophoresis ,Chromatography ,biology ,Chemistry ,Peas ,Membrane Proteins ,biology.organism_classification ,Transmembrane protein ,Plant Leaves ,Transmembrane domain ,Membrane ,Membrane protein - Abstract
The crucial cellular role of membrane proteins is generally known for all life forms. Depending on the species, tissue, compartment, function and physiological condition, membranes differ in their protein and lipid profiles. Additionally, occurrence of microdomains hampers quantitative protein solubilisation and therefore membrane proteomics remain a major challenge. In the present study sample preparation (TCA/acetone and methanol/chloroform precipitation with and without SDS pre-solubilisation) for two-dimensional PAGE were compared for microsomal fractions of leaves (Arabidopsis thaliana, Nicotiana tabaccum, Pisum sativum) and roots (P. sativum, Zea mays). Generally, pre-solubilisation with SDS impaired the resolution of the gels. All samples showed higher spot yields with TCA/acetone precipitation. Finally, we compared the results of conventional 2D-PAGE (IPG/SDS-PAGE) and the combination of off-gel fractionation in the first-dimension, 10% urea-SDS-PAGE in the second-dimension. Results showed that more spots are present in the alkaline pH range after off-gel fractionation then on conventional 2D-PAGE. For the first time, off-gel fractionation was combined with SDS/SDS-PAGE and BAC/SDS-PAGE to improve the resolution after off-gel fractionation. Transmembrane domains and GRAVY were calculated for all significantly identified spots resulting from the MALDI-TOF-TOF mass spectrometry showing that in the second dimension after off-gel fractionation 10.3% more transmembrane proteins were identified compared to IPG/SDS-PAGE.
- Published
- 2011
48. The plasma membrane proteome of maize roots grown under low and high iron conditions
- Author
-
Stefanie Wienkoop, Sabine Lüthje, and David Hopff
- Subjects
Proteomics ,Protein Folding ,Proteome ,Iron ,Biophysics ,Biology ,medicine.disease_cause ,Biochemistry ,Zea mays ,Cell wall ,chemistry.chemical_compound ,Cell Wall ,Heat shock protein ,medicine ,NAD(P)H Dehydrogenase (Quinone) ,Homeostasis ,Nicotianamine ,Heme ,Cytoskeleton ,Plant Proteins ,chemistry.chemical_classification ,Reactive oxygen species ,Cell Membrane ,Oxidative Stress ,chemistry ,Oxidation-Reduction ,Oxidative stress ,Signal Transduction - Abstract
Iron (Fe) homeostasis is essential for life and has been intensively investigated for dicots, while our knowledge for species in the Poaceae is fragmentary. This study presents the first proteome analysis (LC-MS/MS) of plasma membranes isolated from roots of 18-day old maize (Zea mays L.). Plants were grown under low and high Fe conditions in hydroponic culture. In total, 227 proteins were identified in control plants, whereas 204 proteins were identified in Fe deficient plants and 251 proteins in plants grown under high Fe conditions. Proteins were sorted by functional classes, and most of the identified proteins were classified as signaling proteins. A significant number of PM-bound redox proteins could be identified including quinone reductases, heme and copper-containing proteins. Most of these components were constitutive, and others could hint at an involvement of redox signaling and redox homeostasis by change in abundance. Energy metabolism and translation seem to be crucial in Fe homeostasis. The response to Fe deficiency includes proteins involved in development, whereas membrane remodeling and assembly and/or repair of Fe-S clusters is discussed for Fe toxicity. The general stress response appears to involve proteins related to oxidative stress, growth regulation, an increased rigidity and synthesis of cell walls and adaption of nutrient uptake and/or translocation. This article is part of a Special Issue entitled: Plant Proteomics in Europe.
- Published
- 2011
49. Penetration by Artificial Electron Acceptors of the Plasma Membrane-Bound Redox System into Intact Zea mays L. Roots Investigated by Proton-Induced X-Ray Emission
- Author
-
D. GroBmann, Sabine Lüthje, M. Niecke, Michael Böttger, and Olaf Döring
- Subjects
chemistry.chemical_classification ,Physiology ,Chemistry ,Analytical chemistry ,X-ray ,Plant Science ,Penetration (firestop) ,Plasma ,Membrane transport ,Electron acceptor ,Redox ,Apoplast ,Zea mays ,Genetics ,Research Article - Abstract
Proton-induced x-ray emission was used to investigate the penetration of compounds of the membrane-impermeant electron acceptors hexabromoiridate IV, hexachloroiridate IV, and hexacyanoferrate III into corn (Zea mays L.) roots. Maps of the heavy element distribution in cross-sections of fixed, epoxy-embedded roots showed for hexabromoiridate IV small amounts of Br in samples treated for 24 h with concentrations normally used in physiological experiments (0.02 mM). After treatment with high concentrations (0.8 mM) of these complexes, Fe and Ir as well as Br were found in root cross-sections. In samples taken at a distance of 5 mm behind the root tip, we found an even distribution of Fe, Ir, and Br over the whole cross-section. In samples taken 15 mm behind the root tip, about 99% of both Br and Ir was confined to the rhizodermal cell layer. The distribution did not change with the complex used. These data are consistent with the view that apoplastic diffusion of the electron acceptors was blocked by the hypodermal Casparian band.
- Published
- 1993
50. Phylogeny, topology, structure and functions of membrane-bound class III peroxidases in vascular plants
- Author
-
David Hopff, Sabine Lüthje, Claudia-Nicole Meisrimler, and Benjamin Möller
- Subjects
Models, Molecular ,Proteomics ,Protein Conformation ,Plant Science ,Vacuole ,Horticulture ,Biology ,medicine.disease_cause ,Topology ,Biochemistry ,Isozyme ,Phylogenetics ,Plant Cells ,medicine ,Molecular Biology ,Secretory pathway ,Phylogeny ,chemistry.chemical_classification ,Reactive oxygen species ,General Medicine ,Plants ,chemistry ,Peroxidases ,biology.protein ,Oxidative stress ,Peroxidase ,Protein Binding - Abstract
Peroxidases are key player in the detoxification of reactive oxygen species during cellular metabolism and oxidative stress. Membrane-bound isoenzymes have been described for peroxidase superfamilies in plants and animals. Recent studies demonstrated a location of peroxidases of the secretory pathway (class III peroxidases) at the tonoplast and the plasma membrane. Proteomic approaches using highly enriched plasma membrane preparations suggest organisation of these peroxidases in microdomains, a developmentally regulation and an induction of isoenzymes by oxidative stress. Phylogenetic relations, topology, putative structures, and physiological function of membrane-bound class III peroxidases will be discussed.
- Published
- 2010
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