44 results on '"Matzapetakis M"'
Search Results
2. Synthesis, spectroscopic, and structural characterization of the first aqueous cobalt(II)-citrate complex: toward a potentially bioavailable form of cobalt in biologically relevant fluids
- Author
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Matzapetakis, M., Dakanali, M., Raptopoulou, C. P., Tangoulis, V., Terzis, A., Moon, N., Giapintzakis, J., and Salifoglou, A.
- Published
- 2000
- Full Text
- View/download PDF
3. Studies of the Yap8 - DNA interaction: P09-52
- Author
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Amaral, C., Matos, R., Matzapetakis, M., Arraiano, C., and Rodrigues-Pousada, C.
- Published
- 2012
4. Berry fruits modulate kidney dysfunction and urine metabolome in Dahl salt-sensitive rats
- Author
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Gomes, A., primary, Godinho-Pereira, J., additional, Oudot, C., additional, Sequeira, C.O., additional, Macià, A., additional, Carvalho, F., additional, Motilva, M.J., additional, Pereira, S.A., additional, Matzapetakis, M., additional, Brenner, C., additional, and Santos, C.N., additional
- Published
- 2020
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- View/download PDF
5. Active and prospective latent tuberculosis are associated with different metabolomic profiles: clinical potential for the identification of rapid and non-invasive biomarkers
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Albors-Vaquer, A., primary, Rizvi, A., additional, Matzapetakis, M., additional, Lamosa, P., additional, Coelho, A. V., additional, Patel, A. B., additional, Mande, S. C., additional, Gaddam, S., additional, Pineda-Lucena, A., additional, Banerjee, S., additional, and Puchades-Carrasco, L., additional
- Published
- 2020
- Full Text
- View/download PDF
6. Manganese citrate chemistry: syntheses, spectroscopic studies, and structural characterizations of novel mononuclear, water-soluble manganese citrate complexes
- Author
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Matzapetakis, M., Karligiano, N., Bino, A., Dakanali, M., Raptopoulou, C.P., Tangoulis, V., Terzis, A., Giapintzakis, J., and Salifoglou, A.
- Subjects
Manganese in human nutrition -- Research ,Citrates -- Physiological aspects ,Complex compounds -- Spectra ,Bioavailability -- Analysis ,Chemistry - Abstract
Research describes the syntheses, separation, and structural studies of the manganese citrate complexes in aqueous solutions. Data suggest that in these complexes, the manganese ion is bound to two citrate ligands and they exhibit paramagnetic di- and tri-valent manganese species in the solid-state.
- Published
- 2000
7. Lead–citrate chemistry. Synthesis, spectroscopic and structural studies of a novel lead(II)–citrate aqueous complex
- Author
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Kourgiantakis, M., Matzapetakis, M., Raptopoulou, C.P., Terzis, A., and Salifoglou, A.
- Published
- 2000
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8. Solution structure of the cinaciguat bound human beta1 H-NOX.
- Author
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Matzapetakis, M., primary and Saraiva, I.H., additional
- Published
- 2018
- Full Text
- View/download PDF
9. Synthesis, pH-dependent structural characterization, and solution behavior of aqueous aluminium and gallium citrate complexes
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Matzapetakis, M., Salifoglou, A., Kourgiantakis, M., Mavromoustakos, T., Dakanali, M., Iordanidis, L., Raptopoulou, C.P., Banyai, I., Terzis, A., Lakatos, A., and Kiss, T.
- Subjects
Hydrogen-ion concentration -- Analysis ,Gallium compounds -- Chemical properties ,Gallium compounds -- Spectra ,Aluminum compounds -- Chemical properties ,Aluminum compounds -- Spectra ,Chemistry - Abstract
The reactivity of Al(III) and Ga(III) toward citrate ions in aqueous solutions yielded, under stoichiometric and pH-dependent conditions, well-behaved crystalline mononuclear metal-citrate complexes. Aluminium is found to linked with Alzheimer's disease and gallium is found to accumulate in tissues causing tumors.
- Published
- 2001
10. Solution structure human HCN2 CNBD in the cAMP-unbound state
- Author
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Saponaro, A., primary, Pauleta, S.R., additional, Cantini, F., additional, Matzapetakis, M., additional, Hammann, C., additional, Banci, L., additional, Thiel, G., additional, Santoro, B., additional, and Moroni, A., additional
- Published
- 2014
- Full Text
- View/download PDF
11. Synthesis, structural characterization, and solution behavior of the first mononuclear, aqueous aluminum citrate complex
- Author
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Matzapetakis, M., Raptopoulou, C.P., Terzis, A., Lakatos, A., Kiss, T., and Salifoglou, A.
- Subjects
Aluminum compounds -- Research ,Citrates -- Research ,Inorganic compounds -- Synthesis ,Chemistry - Abstract
The synthesis, isolation, structural characterization and solution behavior of the first mononuclear aluminum citrate complex illustrate the significance of the interaction between aluminum and citrate in biological media. The chemical and structural characteristics of the low molecular weight aluminum citrate species help provide insight into citrate's ability to enhance aluminum's absorption and thus impact on its accumulation and biotoxicity at biological sites.
- Published
- 1999
12. Synthesis, spectroscopic and structural characterization of the first mononuclear, water soluble iron-citrate complex, (NH4)5Fe(C6H4O7)2*2H2O
- Author
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Matzapetakis, M., Raptopoulou, C.P., Tsohos, A., Papaefthymiou, V., Moon, N., and Salifoglou, A.
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Nuclear chemistry -- Research ,Iron compounds -- Research ,Citric acid -- Research ,Citrates -- Research ,Chemical reactions -- Research ,Chemistry - Abstract
The aqueous coordination chemistry of iron with citric acid was investigated. Specifically, the synthesis, spectroscopic and structural properties of the first mononuclear, water soluble iron-citrate complex, (NH4)5Fe(C6H4O7)2*2H2O, were described. Reaction between iron(III) nitrate and citric acid with a 1:2 molar ratio in aqueous solution resulted in the isolation of yellow crystalline material consistent with the molecular formula (NH4)5Fe(C6H4O7)2*2H2O.
- Published
- 1998
13. Staphylococcal Nuclease PHS variant
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Matzapetakis, M., primary, Pais, T.M., additional, Lamosa, P., additional, Turner, D.L., additional, and Santos, H., additional
- Published
- 2012
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14. Comparison of the binding of cadmium(II), mercury(II), and arsenic(III) to the de novo designed peptides TRI L12C and TRI L16C
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Matzapetakis, M., Farrer, B. T., Weng, T. C., Hemmingsen, L., Penner-Hahn, J. E., Pecoraro, V. L., Matzapetakis, M., Farrer, B. T., Weng, T. C., Hemmingsen, L., Penner-Hahn, J. E., and Pecoraro, V. L.
- Published
- 2002
15. Synthesis, Spectroscopic and Structural Characterization of the First Mononuclear, Water Soluble Iron−Citrate Complex, (NH4)5Fe(C6H4O7)2·2H2O
- Author
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Matzapetakis, M., primary, Raptopoulou, C. P., additional, Tsohos, A., additional, Papaefthymiou, V., additional, Moon, N., additional, and Salifoglou, A., additional
- Published
- 1998
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16. Structural rearrangements occurring on HCN2 CNBD domain upon cAMP binding
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Saponaro, A., Matzapetakis, M., anna moroni, and Pauleta, S.
17. Urinary proteome and metabolome in dogs (Canis lupus familiaris): The effect of chronic kidney disease
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Francesco Dondi, Katerina Vasylyeva, Elisa Bellei, Manolis Matzapetakis, André M. Almeida, Gloria Isani, Giulia Andreani, Enea Ferlizza, Ferlizza, E, Isani, G, Dondi, F, Andreani, G, Vasylyeva, K, Bellei, E, Almeida, A M, and Matzapetakis, M
- Subjects
0301 basic medicine ,Male ,Tamm–Horsfall protein ,Proteome ,Urinary system ,kidney disease ,Biophysics ,Physiology ,Context (language use) ,Urine ,SDS-PAGE, proteomics, metabolomics, NMR, urine, CKD, dog ,Metabolome ,Dogs ,Chronic kidney disease ,Biochemistry ,03 medical and health sciences ,Metabolomics ,medicine ,Animals ,Renal Insufficiency, Chronic ,Wolves ,030102 biochemistry & molecular biology ,biology ,business.industry ,medicine.disease ,030104 developmental biology ,dog ,biology.protein ,business ,Biomarkers ,Kidney disease - Abstract
Chronic kidney disease (CKD) is a progressive and irreversible disease. Although urine is an ideal biological sample for proteomics and metabolomics studies, sensitive and specific biomarkers are currently lacking in dogs. This study characterised dog urine proteome and metabolome aiming to identify and possibly quantify putative biomarkers of CKD in dogs. Twenty-two healthy dogs and 28 dogs with spontaneous CKD were selected and urine samples were collected. Urinary proteome was separated by SDS-PAGE and analysed by mass spectrometry, while urinary metabolome was analysed in protein-depleted samples by 1D 1H NMR spectra. The most abundant proteins in urine samples from healthy dogs were uromodulin, albumin and, in entire male dogs, arginine esterase. In urine samples from CKD dogs, the concentrations of uromodulin and albumin were significantly lower and higher, respectively, than in healthy dogs. In addition, these samples were characterised by a more complex protein pattern indicating mixed glomerular (protein bands ≥65 kDa) and tubular (protein bands
- Published
- 2020
18. Comparison of the Binding of Cadmium(II), Mercury(II), and Arsenic(III) to the de Novo Designed Peptides TRI L12C and TRI L16C
- Author
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Matzapetakis, M
- Published
- 2002
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19. A Ligand-Free Approach towards Coumarin Analogs via Natural Deep Eutectic Solvent-Mediated Suzuki-Miyaura Coupling.
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Katopodi A, Nikolaou N, Kakokefalou V, Alexandratou E, Matzapetakis M, Zervou M, and Detsi A
- Abstract
A ligand-free approach for the Suzuki-Miyaura cross coupling reaction using Natural Deep Eutectic Solvents (NaDES) towards coumarin analogs is described. A model reaction between the synthetically prepared 3-(4-acetyloxy-phenyl)-6-bromo-4-methyl-coumarin ( 3b ) and phenylboronic acid was performed in five different NaDES as well as in pure glycerol, using two inorganic bases and palladium catalysts. The reaction proceeded smoothly in Choline Chloride/Glycerol (ChCl/Gly) and Betaine/Glycerol (Bet/Gly) NaDES at 90 °C in 24 h, affording the desired product in high yields up to 95%. The combination of K
2 CO3 , Pd(OAc)2 and ChCl/Gly NaDES provided optimum yields and high purity of the desired compounds, while the solvent was successfully recycled and reused up to two times. The developed methodology is applicable to boronic acids bearing various substituents. The formation of palladium nanoparticles in the reaction mixture was observed, and the size of the nanoparticles was associated with the reaction yield. In addition, in all the glycerol-based NaDES, an effective removal of the acetyl group of the acetyloxy-coumarin analogs was observed; thus, it is noteworthy that the Suzuki-Miyaura coupling and the deacetylation reaction were achieved in one pot. The ten novel coumarin derivatives synthesized were structurally characterized using 1D and 2D NMR spectroscopy and were tested for their cytotoxicity against the A431 squamous cancer cell line, presenting significant activity.- Published
- 2024
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20. Goat mammary gland metabolism: An integrated Omics analysis to unravel seasonal weight loss tolerance.
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Ribeiro DM, Palma M, Salvado J, Hernández-Castellano LE, Capote J, Castro N, Argüello A, Matzapetakis M, Araújo SS, and de Almeida AM
- Subjects
- Animals, Female, Seasons, Biomarkers analysis, Weight Loss, Mammary Glands, Animal metabolism, Milk metabolism, Lactation, Goats genetics, Metabolomics
- Abstract
Seasonal weight loss (SWL), is a major limitation to animal production. In the Canary Islands, there are two dairy goat breeds with different levels of tolerance to SWL: Majorera (tolerant) and Palmera (susceptible). Our team has studied the response of these breeds to SWL using different Omics tools. The objective of this study was to integrate such results in a data driven approach and using dedicated tools, namely the DIABLO method. The outputs of our analysis mainly separate unrestricted from restricted goats. Metabolites behave as "hub" molecules, grouping interactions with several genes and proteins. Unrestricted goats upregulated protein synthesis, along with arginine catabolism and adipogenesis pathways, which are related with higher anabolic rates and a larger proportion of secretory tissue, in agreement with their higher milk production. Contrarily, restricted goats seemingly increased the synthesis of acetyl-CoA through serine and acetate conversion into pyruvate. This may have occurred to increase fatty acid synthesis and/or to use them as an energy source in detriment to glucose, which was more available in the diet of unrestricted goats. Lastly, restricted Palmera upregulated the expression of PEBP4 and GPD1 genes compared to all other groups, which might support their use as putative biomarkers for SWL susceptibility. SIGNIFICANCE: Seasonal weight loss (SWL) is a major issue influencing animal production in the tropics and Mediterranean. By studying its impact on the mammary gland of tolerant and susceptible dairy goat breeds, using Omics, we aim at surveying the tissue for possible biomarkers that reflect these traits. In this study, data integration of three Omics (transcriptomics, proteomics and metabolomics) was performed using bioinformatic tools, to relate putative biomarkers and evaluate all three levels of information; in a novel approach. This information can enhance selection programs, lowering the impact of SWL on food production systems., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)
- Published
- 2023
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21. Serum metabolomic profiling unveils distinct sex-related metabolic patterns in NAFLD.
- Author
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Fotakis C, Kalafati IP, Amanatidou AI, Andreou V, Matzapetakis M, Kafyra M, Varlamis I, Zervou M, and Dedoussis GV
- Subjects
- Humans, Female, Male, Metabolomics methods, Obesity metabolism, Metabolome, Non-alcoholic Fatty Liver Disease complications
- Abstract
Objective: Obesity poses an increased risk for the onset of Nonalcoholic fatty liver disease (NAFLD). The influence of other factors, such as sex in the incidence and severity of this liver disease has not yet been fully elucidated. Thus, we aimed to identify the NAFLD serum metabolic signatures associated with sex in normal, overweight and obese patients and to associate the metabolite fluctuations across the increasing liver steatosis stages., Methods and Results: Using nuclear magnetic resonance (NMR) serum samples of 210 NAFLD cases and control individuals diagnosed with liver U/S, our untargeted metabolomics enquiry provided a sex distinct metabolic bouquet. Increased levels of alanine, histidine and tyrosine are associated with severity of NAFLD in both men and women. Moreover, higher serum concentrations of valine, aspartic acid and mannose were positively associated with the progression of NAFLD among the male subjects, while a negative association was observed with the levels of creatine, phosphorylcholine and acetic acid. On the other hand, glucose was positively associated with the progression of NAFLD among the female subjects, while levels of threonine were negatively related. Fluctuations in ketone bodies acetoacetate and acetone were also observed among the female subjects probing a significant reduction in the circulatory levels of the former in NAFLD cases. A complex glycine response to hepatic steatosis of the female subjects deserves further investigation., Conclusion: Results of this study aspire to address the paucity of data on sex differences regarding NAFLD pathogenesis. Targeted circulatory metabolome measurements could be used as diagnostic markers for the distinct stages of NAFLD in each sex and eventually aid in the development of novel sex-related therapeutic options., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Fotakis, Kalafati, Amanatidou, Andreou, Matzapetakis, Kafyra, Varlamis, Zervou and Dedoussis.)
- Published
- 2023
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22. Effect of dietary Spirulina (Arthrospira platensis) on the intestinal function of post-weaned piglet: An approach combining proteomics, metabolomics and histological studies.
- Author
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Martins CF, Ribeiro DM, Matzapetakis M, Pinho MA, Kuleš J, Horvatić A, Guillemin N, Eckersall PD, Freire JPB, de Almeida AM, and Prates JAM
- Subjects
- Animal Feed analysis, Animals, Diet, Dietary Supplements, Glucose, Glutamates, Glycine, Inositol, Lactates, Male, Muramidase, Proteome, Proteomics, Swine, Spirulina
- Abstract
The effect of dietary Spirulina (Arthrospira platensis) and CAZyme supplementation was assessed on the gut of weaned piglets, using an integrated NMR-metabolomics approach combined with Tandem Mass Tag labelled proteomics. Thirty weaned male piglets were assigned to one of the three following diets (n = 10): cereal and soybean meal basal diet (Control), basal diet with 10% Spirulina inclusion (SP) and SP diet supplemented with 0.01% lysozyme (SP + L). The experiment lasted 4 weeks and, upon slaughter, small intestine samples were collected for histological, metabolomic and proteomic analysis. No significant differences were found for the histology and metabolomics analysis between the three experimental groups. Lactate, glutamate, glycine and myo-inositol were the most abundant metabolites. Proteomics results showed 1502 proteins identified in the intestine tissue. A total of 23, 78, 27 differentially abundant proteins were detected respectively for the SP vs. Control, SP + L vs. Control and SP + L vs. SP comparisons. The incorporation of Spirulina and supplementation of lysozyme in the piglet's diets is associated to intestinal proteomic changes. These include increased protein synthesis and abundance of contractile apparatus proteins, related with increased nutrient availability, which has beneficial (increased glucose uptake) and detrimental (increased digesta viscosity) metabolic effects. SIGNIFICANCE: The use of conventional feedstuffs becomes increasingly prohibitive due to its environmental toll. To increase the sustainability of the livestock sector, novel feedstuffs such as microalgae need to be considered. However, its recalcitrant cell wall has antinutritional effects that can inhibit high dietary inclusion levels. The supplementation with CAZymes is a possible solution to this issue. The small intestine is a central piece in monogastric digestion and of particular importance for the weaned piglet. Studying the effect of dietary Spirulina and CAZyme supplementation on its histomorphology, metabolome and proteome allows studying relevant physiological adaptations to these diets., Competing Interests: Declaration of Competing Interest None., (Copyright © 2022 Elsevier B.V. All rights reserved.)
- Published
- 2022
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23. Urinary proteome and metabolome in dogs (Canis lupus familiaris): The effect of chronic kidney disease.
- Author
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Ferlizza E, Isani G, Dondi F, Andreani G, Vasylyeva K, Bellei E, Almeida AM, and Matzapetakis M
- Subjects
- Animals, Biomarkers metabolism, Dogs, Male, Metabolome, Metabolomics, Proteome, Renal Insufficiency, Chronic diagnosis, Wolves metabolism
- Abstract
Chronic kidney disease (CKD) is a progressive and irreversible disease. Although urine is an ideal biological sample for proteomics and metabolomics studies, sensitive and specific biomarkers are currently lacking in dogs. This study characterised dog urine proteome and metabolome aiming to identify and possibly quantify putative biomarkers of CKD in dogs. Twenty-two healthy dogs and 28 dogs with spontaneous CKD were selected and urine samples were collected. Urinary proteome was separated by SDS-PAGE and analysed by mass spectrometry, while urinary metabolome was analysed in protein-depleted samples by 1D
1 H NMR spectra. The most abundant proteins in urine samples from healthy dogs were uromodulin, albumin and, in entire male dogs, arginine esterase. In urine samples from CKD dogs, the concentrations of uromodulin and albumin were significantly lower and higher, respectively, than in healthy dogs. In addition, these samples were characterised by a more complex protein pattern indicating mixed glomerular (protein bands ≥65 kDa) and tubular (protein bands <65 kDa) proteinuria. Urine spectra acquired by NMR allowed the identification of 86 metabolites in healthy dogs, belonging to 49 different pathways mainly involved in amino acid metabolism, purine and aminoacyl-tRNA biosynthesis or tricarboxylic acid cycle. Seventeen metabolites showed significantly different concentrations when comparing healthy and CKD dogs. In particular, carnosine, trigonelline, and cis-aconitate, might be suggested as putative biomarkers of CKD in dogs. SIGNIFICANCE: Urine is an ideal biological sample, however few proteomics and metabolomics studies investigated this fluid in dogs and in the context of CKD (chronic kidney disease). In this research, applying a multi-omics approach, new insights were gained regarding the molecular changes triggered by this disease in canine urinary proteome and metabolome. In particular, the involvement of the tubular component was highlighted, suggesting uromodulin, trigonelline and carnosine as possible biomarkers of CKD in dogs., Competing Interests: Declaration of Competing Interest None, (Copyright © 2020 Elsevier B.V. All rights reserved.)- Published
- 2020
- Full Text
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24. Metabolome and proteome changes in skeletal muscle and blood of pre-weaning calves fed leucine and threonine supplemented diets.
- Author
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Yu K, Matzapetakis M, Horvatić A, Terré M, Bach A, Kuleš J, Yeste N, Gómez N, Arroyo L, Rodríguez-Tomàs E, Peña R, Guillemin N, de Almeida AM, Eckersall PD, and Bassols A
- Subjects
- Animal Feed analysis, Animals, Cattle, Diet, Dietary Supplements, Leucine metabolism, Male, Milk, Muscle, Skeletal metabolism, Threonine metabolism, Weaning, Metabolome, Proteome metabolism
- Abstract
In pre-weaning calves, both leucine and threonine play important roles in growth and muscle metabolism. In this study, metabolomics, proteomics and clinical chemistry were used to assess the effects of leucine and threonine supplementation added to milk replacer on 14 newborn Holstein male calves: 7 were fed a control diet (Ctrl) and 7 were fed the Ctrl diet supplemented with 0.3% leucine and 0.3% threonine (LT) from 5.6 days of age to 53.6 days. At this time, blood and semitendinosus muscle biopsies were collected for analysis. Integrated metabolomics and proteomics showed that branched-chain amino acids (BCAA) degradation and mitochondrial oxidative metabolism (citrate cycle and respiratory chain) were the main activated pathways in muscle because of the supplementation. BCAA derivatives and metabolites related to lipid mobilization showed the major changes. The deleterious effects of activated oxidative phosphorylation were balanced by the upregulation of antioxidant proteins. An increase in protein synthesis was indicated by elevated aminoacyl-tRNA biosynthesis and increased S6 ribosomal protein phosphorylation in skeletal muscle. In conclusion, LT group showed greater BCAA availability and mitochondrial oxidative activity; as the muscle cells undergo greater aerobic metabolism, antioxidant defenses were activated to compensate for possible cell damage. Data are available via ProteomeXchange (PXD016098). SIGNIFICANCE: Leucine and threonine are essential amino acids for the pre-weaning calf, being of high importance for growth. In this study, we found that leucine and threonine supplementation of milk replacer to feed pre-weaning calves led to differences in the proteome, metabolome and clinical chemistry analytes in skeletal muscle and plasma, albeit no differences in productive performance were recorded. This study extends our understanding on the metabolism in dairy calves and helps optimizing their nutritional status., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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25. Skeletal muscle metabolomics and blood biochemistry analysis reveal metabolic changes associated with dietary amino acid supplementation in dairy calves.
- Author
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Yu K, Matzapetakis M, Valent D, Saco Y, De Almeida AM, Terré M, and Bassols A
- Subjects
- Animal Feed analysis, Animals, Body Weight drug effects, Cattle, Male, Amino Acids pharmacology, Blood Chemical Analysis, Dairying, Dietary Supplements analysis, Metabolomics, Muscle, Skeletal drug effects, Muscle, Skeletal metabolism
- Abstract
The effects of different amino acid (AA) supplementations of milk protein-based milk replacers in pre-ruminant calves from 3 days to 7 weeks of age were studied. Animals were divided into 4 groups: Ctrl) Control group fed with milk protein-based milk replacer without supplementation; GP) supplementation with 0.1% glycine and 0.3% proline; FY) supplementation with 0.2% phenylalanine and 0.2% tyrosine; MKT) supplementation with 0.62% lysine, 0.22% methionine and 0.61% threonine. For statistical analysis, t-test was used to compare AA-supplemented animals to the Ctrl group. At week 7, body weight and average daily gain (ADG) were measured and blood samples and skeletal muscle biopsies were taken. Blood biochemistry analytes related to energy metabolism were determined and it was shown that MKT group had higher serum creatinine and higher plasma concentration of three supplemented AAs as well as arginine compared with the Ctrl group. GP group had similar glycine/proline plasma concentration compared with the other groups while in FY group only plasma phenylalanine concentration was higher compared with Control. Although the AA supplementations in the GP and FY groups did not affect average daily gain and metabolic health profile from serum, the metabolome analysis from skeletal muscle biopsy revealed several differences between the GP-FY groups and the Ctrl-MKT groups, suggesting a metabolic adaptation especially in GP and FY groups.
- Published
- 2018
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26. Mammary gland and milk fatty acid composition of two dairy goat breeds under feed-restriction.
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Palma M, Alves SP, Hernández-Castellano LE, Capote J, Castro N, Argüello A, Matzapetakis M, Bessa RJB, and de Almeida AM
- Subjects
- Animal Feed supply & distribution, Animals, Breeding, Female, Mammary Glands, Animal physiology, Oleic Acid analysis, Palmitic Acid analysis, Spain, Species Specificity, Fatty Acids analysis, Goats physiology, Mammary Glands, Animal chemistry, Milk chemistry, Seasons, Weight Loss
- Abstract
Goat dairy products are an important source of animal protein in the tropics. During the dry season, pasture scarcity leads animals to lose up to 40% of their body weight, a condition known as Seasonal Weight Loss (SWL) that is one of the major constraints in ruminant production. Breeds with high tolerance to SWL are relevant to understand the physiological responses to pasture scarcity so they could be used in programs for animal breeding. In the Canary Islands there are two dairy goat breeds with different levels of tolerance to SWL: the Palmera, susceptible to SWL; and the Majorera, tolerant to SWL. Fat is one of the milk components most affected by environmental and physiological conditions. This study hypothesises that feed-restriction affects Majorera and Palmera breeds differently, leading to different fatty acid profiles in the mammary gland and milk. An interaction between breed and feed-restriction was observed in the mammary gland. Feed-restriction was associated with an increase in oleic acid and a decrease in palmitic acid percentage in the Palmera breed whereas no differences were observed in the Majorera breed. Palmitic and oleic acids together constituted around 60% of the total fatty acids identified, which suggests that Palmera breed is more susceptible to SWL. In milk, feed-restriction affected both breeds similarly. Regarding the interaction of the breed with the treatment, we also observed similar responses in both breeds, but this influence affects only around 2% of the total fatty acids. In general, Majorera breed is more tolerant to feed-restriction.
- Published
- 2017
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27. The hepatic and skeletal muscle ovine metabolomes as affected by weight loss: a study in three sheep breeds using NMR-metabolomics.
- Author
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Palma M, Scanlon T, Kilminster T, Milton J, Oldham C, Greeff J, Matzapetakis M, and Almeida AM
- Subjects
- Adaptation, Physiological, Animals, Breeding, Proton Magnetic Resonance Spectroscopy methods, Seasons, Sheep, Sheep, Domestic, Liver chemistry, Metabolomics methods, Muscle, Skeletal chemistry, Weight Loss
- Abstract
Sheep are a valuable resource for meat and wool production. During the dry summer, pastures are scarce and animals face Seasonal Weight Loss (SWL), which decreases production yields. The study of breeds tolerant to SWL is important to understand the physiological mechanisms of tolerance to nutritional scarcity, and define breeding strategies. Merino, Damara and Dorper sheep breeds have been described as having different levels of tolerance to SWL. In this work, we assess their liver and muscle metabolomes, and compare the responses to feed restriction. Ram lambs from each breed were divided into growth and feed restricted groups, over 42 days. Tissue metabolomes were assessed by
1 H-NMR. The Dorper restricted group showed few changes in both tissues, suggesting higher tolerance to nutritional scarcity. The Merinos exhibited more differences between treatment groups. Major differences were related to fat and protein mobilization, and antioxidant activity. Between the Damara groups, the main differences were observed in amino acid composition in muscle and in energy-related pathways in the liver. Integration of present results and previous data on the same animals support the hypothesis that, Dorper and Damara breeds are more tolerant to SWL conditions and thus, more suitable breeds for harsh environmental conditions.- Published
- 2016
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28. NMR-metabolomics profiling of mammary gland secretory tissue and milk serum in two goat breeds with different levels of tolerance to seasonal weight loss.
- Author
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Palma M, Hernández-Castellano LE, Castro N, Arguëllo A, Capote J, Matzapetakis M, and de Almeida AM
- Subjects
- Adaptation, Biological, Animals, Breeding, Female, Goats, Mammary Glands, Animal metabolism, Metabolome, Metabolomics methods, Milk metabolism, Nuclear Magnetic Resonance, Biomolecular, Seasons, Weight Loss
- Abstract
Goats are of special importance in the Mediterranean and tropical regions for producing a variety of dairy products. The scarcity of pastures during the dry season leads to seasonal weight loss (SWL), which affects milk production. In this work, we studied the effect of feed-restriction on two dairy goat breeds, with different tolerance levels to SWL: the Majorera breed (tolerant) and the Palmera breed (susceptible). Nuclear magnetic resonance (NMR) was used to compare the metabolome of an aqueous fraction of the mammary gland and milk serum from both breeds. Goats in mid-lactation were divided by breed, and each in two feed-regime groups: the control group and the restricted-fed group (to achieve 15-20% reduction of body weight at the end of the experiment). Milk and mammary gland samples were collected at the end of the experimental period (23rd day). (1)H NMR spectra were collected from the aqueous extract of the mammary gland biopsies and the milk serum. Profiling analysis has led to the identification of 46 metabolites in the aqueous extract of the mammary gland. Lactose, glutamate, glycine and lactate were found to be the most abundant. Analysis of milk serum allowed the identification of 50 metabolites, the most abundant being lactose, citrate and creatine. Significant differences were observed, in mammary gland biopsies and milk serum, between control and restricted-fed groups in both breeds, albeit with no differences between the breeds. Variations seem to be related to metabolism adaptation to the low-energy diet and are indicative of breed-specific microflora. Milk serum showed more metabolites varying between control and restricted groups, than the mammary gland. The Majorera breed also showed more variations than the Palmera breed in milk samples, which could be an indication of a prompt adaptation to SWL by the Majorera breed.
- Published
- 2016
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29. The solution structure of the soluble form of the lipid-modified azurin from Neisseria gonorrhoeae, the electron donor of cytochrome c peroxidase.
- Author
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Nóbrega CS, Saraiva IH, Carreira C, Devreese B, Matzapetakis M, and Pauleta SR
- Subjects
- Amino Acid Sequence, Azurin genetics, Azurin metabolism, Cloning, Molecular, Copper metabolism, Cytochrome-c Peroxidase genetics, Cytochrome-c Peroxidase metabolism, Electron Transport, Escherichia coli genetics, Escherichia coli metabolism, Gene Expression, Models, Molecular, Molecular Sequence Data, Neisseria gonorrhoeae enzymology, Oxidation-Reduction, Protein Folding, Protein Structure, Secondary, Protein Structure, Tertiary, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Substrate Specificity, Azurin chemistry, Copper chemistry, Cytochrome-c Peroxidase chemistry, Electrons, Hydrogen Peroxide chemistry, Neisseria gonorrhoeae chemistry
- Abstract
Neisseria gonorrhoeae colonizes the genitourinary track, and in these environments, especially in the female host, the bacteria are subjected to low levels of oxygen, and reactive oxygen and nitrosyl species. Here, the biochemical characterization of N. gonorrhoeae Laz is presented, as well as, the solution structure of its soluble domain determined by NMR. N. gonorrhoeae Laz is a type 1 copper protein of the azurin-family based on its spectroscopic properties and structure, with a redox potential of 277±5 mV, at pH7.0, that behaves as a monomer in solution. The globular Laz soluble domain adopts the Greek-key motif, with the copper center located at one end of the β-barrel coordinated by Gly48, His49, Cys113, His118 and Met122, in a distorted trigonal geometry. The edge of the His118 imidazole ring is water exposed, in a surface that is proposed to be involved in the interaction with its redox partners. The heterologously expressed Laz was shown to be a competent electron donor to N. gonorrhoeae cytochrome c peroxidase. This is an evidence for its involvement in the mechanism of protection against hydrogen peroxide generated by neighboring lactobacilli in the host environment., (Copyright © 2015 Elsevier B.V. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
30. Chemical shift assignments and secondary structure determination of the ectodomain of Bacillus subtilis morphogenic protein RodZ.
- Author
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Pereira AC, Paiva A, Saraiva IH, Costa T, Henriques AO, and Matzapetakis M
- Subjects
- Protein Structure, Secondary, Protein Structure, Tertiary, Proton Magnetic Resonance Spectroscopy, Bacillus subtilis metabolism, Bacterial Proteins chemistry, Nuclear Magnetic Resonance, Biomolecular
- Abstract
RodZ (also known as YfgA) is a component of the core bacterial morphogenic apparatus. RodZ is a key cell shape determinant in rod-shaped bacteria and it interacts with the actin-like cytoskeletal protein MreB. In Bacillus subtilis, this 304-residue transmembrane protein is composed of three distinct domains: a cytoplasmic domain (RodZn), a transmembrane domain, and an extra-cytoplasmic domain (RodZc). Here we report the (1)H, (13)C and (15)N backbone and side chain resonance assignments of the RodZc domain from B. subtilis by NMR spectroscopy, and the resulting secondary structure prediction.
- Published
- 2015
- Full Text
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31. Structural basis for the mutual antagonism of cAMP and TRIP8b in regulating HCN channel function.
- Author
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Saponaro A, Pauleta SR, Cantini F, Matzapetakis M, Hammann C, Donadoni C, Hu L, Thiel G, Banci L, Santoro B, and Moroni A
- Subjects
- Binding Sites, Crystallography, X-Ray, Cyclic AMP metabolism, Cyclic Nucleotide-Gated Cation Channels chemistry, Cyclic Nucleotide-Gated Cation Channels metabolism, Electrophoresis, Polyacrylamide Gel, Humans, Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels metabolism, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Structure, Potassium Channels chemistry, Potassium Channels metabolism, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Receptors, Cytoplasmic and Nuclear metabolism, Cyclic AMP chemistry, Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels chemistry, Ion Channel Gating, Receptors, Cytoplasmic and Nuclear chemistry
- Abstract
cAMP signaling in the brain mediates several higher order neural processes. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels directly bind cAMP through their cytoplasmic cyclic nucleotide binding domain (CNBD), thus playing a unique role in brain function. Neuronal HCN channels are also regulated by tetratricopeptide repeat-containing Rab8b interacting protein (TRIP8b), an auxiliary subunit that antagonizes the effects of cAMP by interacting with the channel CNBD. To unravel the molecular mechanisms underlying the dual regulation of HCN channel activity by cAMP/TRIP8b, we determined the NMR solution structure of the HCN2 channel CNBD in the cAMP-free form and mapped on it the TRIP8b interaction site. We reconstruct here the full conformational changes induced by cAMP binding to the HCN channel CNBD. Our results show that TRIP8b does not compete with cAMP for the same binding region; rather, it exerts its inhibitory action through an allosteric mechanism, preventing the cAMP-induced conformational changes in the HCN channel CNBD.
- Published
- 2014
- Full Text
- View/download PDF
32. Two residues in the basic region of the yeast transcription factor Yap8 are crucial for its DNA-binding specificity.
- Author
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Amaral C, Pimentel C, Matos RG, Arraiano CM, Matzapetakis M, and Rodrigues-Pousada C
- Subjects
- Amino Acid Sequence, Basic-Leucine Zipper Transcription Factors genetics, Conserved Sequence, DNA-Binding Proteins chemistry, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Models, Molecular, Molecular Sequence Data, Pancreatitis-Associated Proteins, Protein Binding, Response Elements, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins genetics, Schizosaccharomyces genetics, Schizosaccharomyces metabolism, Sequence Homology, Transcription Factors genetics, Basic-Leucine Zipper Transcription Factors chemistry, Basic-Leucine Zipper Transcription Factors metabolism, DNA metabolism, Protein Interaction Domains and Motifs genetics, Saccharomyces cerevisiae Proteins chemistry, Saccharomyces cerevisiae Proteins metabolism
- Abstract
In Saccharomyces cerevisiae, the transcription factor Yap8 is a key determinant in arsenic stress response. Contrary to Yap1, another basic region-leucine zipper (bZIP) yeast regulator, Yap8 has a very restricted DNA-binding specificity and only orchestrates the expression of ACR2 and ACR3 genes. In the DNA-binding basic region, Yap8 has three distinct amino acids residues, Leu26, Ser29 and Asn31, at sites of highly conserved positions in the other Yap family of transcriptional regulators and Pap1 of Schizosaccharomyces pombe. To evaluate whether these residues are relevant to Yap8 specificity, we first built a homology model of the complex Yap8bZIP-DNA based on Pap1-DNA crystal structure. Several Yap8 mutants were then generated in order to confirm the contribution of the residues predicted to interact with DNA. Using bioinformatics analysis together with in vivo and in vitro approaches, we have identified several conserved residues critical for Yap8-DNA binding. Moreover, our data suggest that Leu26 is required for Yap8 binding to DNA and that this residue together with Asn31, hinder Yap1 response element recognition by Yap8, thus narrowing its DNA-binding specificity. Furthermore our results point to a role of these two amino acids in the stability of the Yap8-DNA complex.
- Published
- 2013
- Full Text
- View/download PDF
33. ¹H, ¹³C and ¹⁵N resonance assignment of the soluble form of the lipid-modified Azurin from Neisseria gonorrhoeae.
- Author
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Nóbrega CS, Matzapetakis M, and Pauleta SR
- Subjects
- Amino Acid Sequence, Carbon Isotopes, Molecular Sequence Data, Nitrogen Isotopes, Protein Structure, Secondary, Solubility, Azurin chemistry, Lipids chemistry, Neisseria gonorrhoeae metabolism, Nuclear Magnetic Resonance, Biomolecular, Protons
- Abstract
Lipid-modified azurin (Laz) from Neisseria gonorrhoeae is a type 1 copper protein proposed to be the electron donor to several enzymes involved in the resistance mechanism to reactive oxygen and nitrogen species. Here we report the backbone and side-chain resonance assignment of Laz in the reduced form, which has been complete at 97%. The predicted secondary structure indicates that this protein belongs to the azurin subfamily of type 1 copper proteins.
- Published
- 2013
- Full Text
- View/download PDF
34. Computational protein design with electrostatic focusing: experimental characterization of a conditionally folded helical domain with a reduced amino acid alphabet.
- Author
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Suárez-Diez M, Pujol AM, Matzapetakis M, Jaramillo A, and Iranzo O
- Subjects
- Algorithms, Amino Acid Sequence, Circular Dichroism, Computer Simulation, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Sequence Data, Protein Conformation, Protein Folding, Static Electricity, Thermodynamics, Amino Acids chemistry, Computational Biology methods, Protein Engineering methods, Proteins chemistry
- Abstract
Automated methodologies to design synthetic proteins from first principles use energy computations to estimate the ability of the sequences to adopt a targeted structure. This approach is still far from systematically producing native-like sequences, due, most likely, to inaccuracies when modeling the interactions between the protein and its aqueous environment. This is particularly challenging when engineering small protein domains (with less polar pair interactions than with the solvent). We have re-designed a three-helix bundle, domain B, using a fixed backbone and a four amino acid alphabet. We have enlarged the rotamer library with conformers that increase the weight of electrostatic interactions within the design process without altering the energy function used to compute the folding free energy. Our synthetic sequences show less than 15% similarity to any Swissprot sequence. We have characterized our sequences in different solvents using circular dichroism and nuclear magnetic resonance. The targeted structure achieved is dependent on the solvent used. This method can be readily extended to larger domains. Our method will be useful for the engineering of proteins that become active only in a given solvent and for designing proteins in the context of hydrophobic solvents, an important fraction of the situations in the cell., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2013
- Full Text
- View/download PDF
35. Structural characterization by NMR of a double phosphorylated chimeric peptide vaccine for treatment of Alzheimer's disease.
- Author
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Ramírez-Gualito K, Richter M, Matzapetakis M, Singer D, and Berger S
- Subjects
- Acyltransferases chemical synthesis, Acyltransferases immunology, Alzheimer Disease immunology, Alzheimer Disease therapy, Antigens, Bacterial immunology, Bacterial Proteins chemical synthesis, Bacterial Proteins immunology, Humans, Mycobacterium tuberculosis chemistry, Nuclear Magnetic Resonance, Biomolecular, Peptides chemical synthesis, Peptides immunology, Phosphorylation, Protein Structure, Secondary, Vaccines chemical synthesis, Vaccines immunology, tau Proteins chemical synthesis, tau Proteins immunology, Acyltransferases chemistry, Antigens, Bacterial chemistry, Bacterial Proteins chemistry, Peptides chemistry, Vaccines chemistry, tau Proteins chemistry
- Abstract
Rational design of peptide vaccines becomes important for the treatment of some diseases such as Alzheimer's disease (AD) and related disorders. In this study, as part of a larger effort to explore correlations of structure and activity, we attempt to characterize the doubly phosphorylated chimeric peptide vaccine targeting a hyperphosphorylated epitope of the Tau protein. The 28-mer linear chimeric peptide consists of the double phosphorylated B cell epitope Tau₂₂₉₋₂₃₇[pThr231/pSer235] and the immunomodulatory T cell epitope Ag85B₂₄₁₋₂₅₅ originating from the well-known antigen Ag85B of the Mycobacterium tuberculosis, linked by a four amino acid sequence -GPSL-. NMR chemical shift analysis of our construct demonstrated that the synthesized peptide is essentially unfolded with a tendency to form a β-turn due to the linker. In conclusion, the -GPSL- unit presumably connects the two parts of the vaccine without transferring any structural information from one part to the other. Therefore, the double phosphorylated epitope of the Tau peptide is flexible and accessible.
- Published
- 2013
- Full Text
- View/download PDF
36. Mannosylglycerate stabilizes staphylococcal nuclease with restriction of slow β-sheet motions.
- Author
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Pais TM, Lamosa P, Matzapetakis M, Turner DL, and Santos H
- Subjects
- Mannose metabolism, Models, Molecular, Molecular Dynamics Simulation, Protein Denaturation, Protein Folding, Protein Stability, Protein Structure, Secondary, Staphylococcus aureus chemistry, Staphylococcus aureus metabolism, Thermodynamics, Excipients metabolism, Glyceric Acids metabolism, Mannose analogs & derivatives, Micrococcal Nuclease chemistry, Micrococcal Nuclease metabolism, Staphylococcus aureus enzymology
- Abstract
Mannosylglycerate is a compatible solute typical of thermophilic marine microorganisms that has a remarkable ability to protect proteins from thermal denaturation. This ionic solute appears to be a universal stabilizing agent, but the extent of protection depends on the specific protein examined. To understand how mannosylglycerate confers protection, we have been studying its influence on the internal motions of a hyperstable staphylococcal nuclease (SNase). Previously, we found a correlation between the magnitude of protein stabilization and the restriction of fast backbone motions. We now report the effect of mannosylglycerate on the fast motions of side-chains and on the slower unfolding motions of the protein. Side-chain motions were assessed by (13)CH(3) relaxation measurements and model-free analysis while slower unfolding motions were probed by H/D exchange measurements at increasing concentrations of urea. Side-chain motions were little affected by the presence of different concentrations of mannosylglycerate or even by the presence of urea (0.25M), and show no correlation with changes in the thermodynamic stability of SNase. Native hydrogen exchange experiments showed that, contrary to reports on other stabilizing solutes, mannosylglycerate restricts local motions in addition to the global motions of the protein. The protein unfolding/folding pathway remained undisturbed in the presence of mannosylglycerate but the solute showed a specific effect on the local motions of β-sheet residues. This work reinforces the link between solute-induced stabilization and restriction of protein motions at different timescales, and shows that the solute preferentially affects specific structural elements of SNase., (Copyright © 2012 The Protein Society.)
- Published
- 2012
- Full Text
- View/download PDF
37. Backbone and side chain 1H, 15N and 13C assignments for a thiol-disulphide oxidoreductase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125.
- Author
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Collins T, Matzapetakis M, and Santos H
- Subjects
- Antarctic Regions, Carbon Isotopes, Hydrogen, Nitrogen Isotopes, Protein Structure, Secondary, Temperature, Nuclear Magnetic Resonance, Biomolecular, Protein Disulfide Reductase (Glutathione) chemistry, Pseudoalteromonas enzymology
- Abstract
Enzymes produced by psychrophilic organisms have successfully overcome the low temperature challenge and evolved to maintain high catalytic rates in their permanently cold environments. As an initial step in our attempt to elucidate the cold-adaptation strategies used by these enzymes we report here the (1)H, (15)N and (13)C assignments for the reduced form of a thiol-disulphide oxidoreductase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125.
- Published
- 2010
- Full Text
- View/download PDF
38. A method for C(alpha) direct-detection in protonless NMR.
- Author
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Bermel W, Bertini I, Felli IC, Matzapetakis M, Pierattelli R, Theil EC, and Turano P
- Subjects
- Animals, Carbon Isotopes, Nitrogen Isotopes, Rana catesbeiana, Ferritins chemistry, Nuclear Magnetic Resonance, Biomolecular methods, Peptides, Cyclic chemistry, Superoxide Dismutase chemistry
- Abstract
Attempts are made to efficiently decouple (13)C nuclei without significant loss of coherence during the application of the decoupling package. Such attempts are based on the S(3)E spin-state selection method. A newly developed double S(3)E (DS(3)E) is particularly efficient for C(alpha) detection for proteins as large as 480 kDa.
- Published
- 2007
- Full Text
- View/download PDF
39. 13C- 13C NOESY spectra of a 480 kDa protein: solution NMR of ferritin.
- Author
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Matzapetakis M, Turano P, Theil EC, and Bertini I
- Subjects
- Amino Acids chemistry, Animals, Carbon chemistry, Carbon Isotopes, Molecular Weight, Protein Structure, Secondary, Rana catesbeiana, Solutions, Time Factors, Carbon analysis, Ferritins chemistry, Nuclear Magnetic Resonance, Biomolecular methods
- Abstract
Molecular size has limited solution NMR analyses of proteins. We report (13)C-(13)C NOESY experiments on a 480 kDa protein, the multi-subunit ferritin nanocage with gated pores. By exploiting (13)C-resonance-specific chemical shifts and spin diffusion effects, we identified 75% of the amino acids, with intraresidue C-C connectivities between nuclei separated by 1-4 bonds. These results show the potential of (13)C-(13)C NOESY for solution studies of molecular assemblies >100 kDa.
- Published
- 2007
- Full Text
- View/download PDF
40. Peptidic models for the binding of Pb(II), Bi(III) and Cd(II) to mononuclear thiolate binding sites.
- Author
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Matzapetakis M, Ghosh D, Weng TC, Penner-Hahn JE, and Pecoraro VL
- Subjects
- Amino Acid Sequence, Binding Sites, Bismuth chemistry, Cadmium chemistry, Circular Dichroism, Lead chemistry, Metalloproteins metabolism, Metals, Heavy metabolism, Molecular Sequence Data, Cysteine chemistry, Metalloproteins chemistry, Metals, Heavy chemistry, Models, Chemical, Peptides chemistry
- Abstract
Herein, we evaluate the binding of Pb(II) and Bi(III) to cysteine-substituted versions of the TRI peptides [AcG-(LKALEEK)4G-NH2] which have previously been shown to bind Hg(II) and Cd(II) in unusual geometries as compared with small-molecule thiol ligands in aqueous solutions. Studies of Pb(II) and Bi(III) with the peptides give rise to complexes consistent with the metal ions bound to three sulfur atoms with M-S distances of 2.63 and 2.54 A, respectively. Competition experiments between the metal ions Pb(II), Cd(II), Hg(II) and Bi(III) for the peptides show that Hg(II) has the highest affinity, owing to the initial formation of the extremely strong HgS2 bond. Cd(II) and Pb(II) have comparable binding affinities at pH > 8, while Bi(III) displays the weakest affinity, following the model, M(II) + (TRI LXC)3(3-) --> M(II)(TRI LXC)3(-). While the relevant equilibria for Hg(II) binding to the TRI peptides corresponds to a strong first step forming Hg(TRI LXC)2(HTRI LXC), followed by a single deprotonation to give Hg(TRI LXC)3(-), the binding of Cd(II) and Pb(II) is consistent with initial formation of M(II)(TRI LXC)(HTRI LXC)2 (+) at pH < 5 followed by a two-proton dissociation step (pK(a2)) yielding M(II)(TRI LXC)3(-). Pb(II)(TRI LXC)(HTRI LXC)2(+) converts to Pb(II)(TRI LXC)3(-) at slightly lower pH values than the corresponding Cd(II)-peptide complexes. In addition, Pb(II) displays a lower pK (a) of binding to the "d"-substituted peptide, (TRI L12C, pK(a2) = 12.0) compared with the "a"-substituted peptide, (TRI L16C, pK (a2) = 12.6), the reverse of the order seen for Hg(II) and Cd(II). Pb(II) also showed a stronger binding affinity for TRI L12C (K(bind) = 3.2 x 10(7) M(-1)) compared with that with TRI L16C (K(bind) = 1.2 x 10(7) M(-1)) at pH > 8.
- Published
- 2006
- Full Text
- View/download PDF
41. Ferritins: iron/oxygen biominerals in protein nanocages.
- Author
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Theil EC, Matzapetakis M, and Liu X
- Subjects
- Catalysis, Ferritins biosynthesis, Ferritins genetics, Gene Expression Regulation, Ferritins metabolism, Iron chemistry, Nanostructures chemistry, Oxygen chemistry, Proteins chemistry
- Abstract
Ferritin protein nanocages that form iron oxy biominerals in the central nanometer cavity are nature's answer to managing iron and oxygen; gene deletions are lethal in mammals and render bacteria more vulnerable to host release of antipathogen oxidants. The multifunctional, multisubunit proteins couple iron with oxygen (maxi-ferritins) or hydrogen peroxide (mini-ferritins) at catalytic sites that are related to di-iron sites oxidases, ribonucleotide reductase, methane monooxygenase and fatty acid desaturases, and synthesize mineral precursors. Gated pores, distributed symmetrically around the ferritin cages, control removal of iron by reductants and chelators. Gene regulation of ferritin, long known to depend on iron and, in animals, on a noncoding messenger RNA (mRNA) structure linked in a combinatorial array to functionally related mRNA of iron transport, has recently been shown to be linked to an array of proteins for antioxidant responses such as thioredoxin and quinone reductases. Ferritin DNA responds more to oxygen signals, and ferritin mRNA responds more to iron signals. Ferritin genes (DNA and RNA) and protein function at the intersection of iron and oxygen chemistry in biology.
- Published
- 2006
- Full Text
- View/download PDF
42. Site-selective metal binding by designed alpha-helical peptides.
- Author
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Matzapetakis M and Pecoraro VL
- Subjects
- Amino Acid Sequence, Binding Sites, Cadmium metabolism, Metalloproteins chemical synthesis, Metalloproteins metabolism, Molecular Sequence Data, Nuclear Magnetic Resonance, Biomolecular, Peptides chemical synthesis, Peptides metabolism, Protein Structure, Secondary, Cadmium chemistry, Metalloproteins chemistry, Peptides chemistry
- Abstract
It is known that the designed alpha-helical peptide family TRI [(Ac-G(LKALEEK)4G-CONH2)], containing single site substitution of a cysteine for a leucine, is capable of binding Cd(II) within a three-stranded coiled coil. The binding affinity of cadmium is dependent upon the site of substitution, with cysteine incorporated at the a site leading to cadmium complexes of higher affinity than when a d site was modified. In this work we have examined whether this differential binding affinity can be expressed in a di-cysteine-substituted peptide in order to develop site specificity within a designed system. The peptide TRI L9CL19C was used to determine whether significant differences in binding affinities at nearly proximal sites could be achieved in a short designed peptide. On the basis of 113Cd, 1H NMR, and circular dichroic spectroscopies, we have shown that 1 equiv of Cd(II) binds exclusively at the a site. Only after that position is filled does the second site become populated. Thus, the TRI system represents the first example where stoichiometrically equivalent peptides with different sequences form the framework for designing molecular assemblies that show site-specific ion recognition. We propose that the distinct metal affinities are due to the cysteine conformers at different substitution points along the peptide. Furthermore, we have shown that site selectivity in biomolecules can be encoded into relatively short peptides with helical sequences and, therefore, do not necessarily require the extensive protein scaffolds found in natural systems.
- Published
- 2005
- Full Text
- View/download PDF
43. Control of metal coordination number in de novo designed peptides through subtle sequence modifications.
- Author
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Lee KH, Matzapetakis M, Mitra S, Marsh EN, and Pecoraro VL
- Subjects
- Amino Acid Sequence, Hydrophobic and Hydrophilic Interactions, Leucine chemistry, Metalloproteins chemical synthesis, Molecular Sequence Data, Nuclear Magnetic Resonance, Biomolecular, Cadmium chemistry, Metalloproteins chemistry, Peptides chemistry
- Abstract
Substitution of an alanine for leucine (shown in light blue) in the hydrophobic interior of designed three-stranded coiled coils allows for the control of metal ion coordination number and geometry. The influence of this perturbation by a noncoordinating residue can be monitored by the dramatic impact on the 113Cd NMR spectrum. The structural effect occurs even when the residue substitution is as much as 7 A from the metal binding site.
- Published
- 2004
- Full Text
- View/download PDF
44. Comparison of the binding of cadmium(II), mercury(II), and arsenic(III) to the de novo designed peptides TRI L12C and TRI L16C.
- Author
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Matzapetakis M, Farrer BT, Weng TC, Hemmingsen L, Penner-Hahn JE, and Pecoraro VL
- Subjects
- Amino Acid Sequence, Arsenic chemistry, Circular Dichroism, Kinetics, Mercury chemistry, Metalloproteins chemical synthesis, Molecular Sequence Data, Spectrophotometry, Ultraviolet, Cadmium chemistry, Metalloproteins chemistry
- Abstract
Designed alpha-helical peptides of the TRI family with a general sequence Ac-G(LKALEEK)(4)G-CONH(2) were used as model systems for the study of metal-protein interactions. Variants containing cysteine residues in positions 12 (TRI L12C) and 16 (TRI L16C) were used for the metal binding studies. Cd(II) binding was investigated, and the results were compared with previous and current work on Hg(II) and As(III) binding. The metal peptide assemblies were studied with the use of UV, CD, EXAFS, (113)Cd NMR, and (111m)Cd perturbed angular correlation spectroscopy. The metalated peptide aggregates exhibited pH-dependent behavior. At high pH values, Cd(II) was bound to the three sulfurs of the three-stranded alpha-helical coiled coils. A mixture of two species was observed, including Cd(II) in a trigonal planar geometry. The complexes have UV bands at 231 nm (20 600 M(-1) cm(-1)) for TRI L12C and 232 nm (22 600 M(-1) cm(-1)) for TRI L16C, an average Cd-S bond length of 2.49 A for both cases, and a (113)Cd NMR chemical shift at 619 ppm (Cd(II)(TRI L12C)(3)(-)) or 625 ppm (Cd(II)(TRI-L16C)(3)(-)). Nuclear quadrupole interactions show that two different Cd species are present for both peptides. One species with omega(0) = 0.45 rad/ns and low eta is attributed to a trigonal planar Cd-(Cys)(3) site. The other, with a smaller omega(0), is attributed to a four-coordinate Cd(Cys)(3)(H(2)O) species. At low pH, no metal binding was observed. Hg(II) binding to TRI L12C was also found to be pH dependent, and a 3:1 sulfur-to-mercury(II) species was observed at pH 9.4. These metal peptide complexes provide insight into heavy metal binding and metalloregulatory proteins such as MerR or CadC.
- Published
- 2002
- Full Text
- View/download PDF
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