131 results on '"Sarg, Bettina"'
Search Results
102. The Microheterogeneity of the Mammalian H10Histone
- Author
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Lindner, Herbert, primary, Sarg, Bettina, additional, Hoertnagl, Brigitte, additional, and Helliger, Wilfried, additional
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- 1998
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103. Substrate and sequential site specificity of cytoplasmic histone acetyltransferases of maize and rat liver
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Kölle, Doris, primary, Sarg, Bettina, additional, Lindner, Herbert, additional, and Loidl, Peter, additional
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- 1998
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104. Application of hydrophilic-interaction liquid chromatography to the separation of phosphorylated H1 histones
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Lindner, Herbert, primary, Sarg, Bettina, additional, and Helliger, Wilfried, additional
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- 1997
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105. Separation of acetylated core histones by hydrophilic-interaction liquid chromatography
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Lindner, Herbert, primary, Sarg, Bettina, additional, Meraner, Christoph, additional, and Helliger, Wilfried, additional
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- 1996
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106. Effect of buffer composition on the migration order and separation of histone H1 subtypes
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Lindner, Herbert, primary, Helliger, Wilfried, additional, Sarg, Bettina, additional, and Meraner, Christoph, additional
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- 1995
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107. Histone H5-chromatin interactions in situ are strongly modulated by H5 C-terminal phosphorylation.
- Author
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Kostova, Nora N., Srebreva, Ljuba, Markov, Dimiter V., Sarg, Bettina, Lindner, Herbert H., and Rundquist, Ingemar
- Abstract
We used linker histone-depleted normal human fibroblast nuclei as templates to study how phosphorylation affects histone H5 binding to chromatin in situ. Permeabilized cells were treated with 0.7 M NaCl to extract the native linker histones. Histone H5 was purified from chicken erythrocytes and phosphorylated in vitro by recombinant cdk5/p35 kinase. High performance capillary electrophoresis (HPCE) showed that the phosphorylated protein contained a mixture of multiply phosphorylated forms. Control experiments, using mass spectrometry, revealed that up to five SPXK motifs in the C terminus were phosphorylated, but also that about 10% of the protein contained one phosphoserine in the N-terminus. Reconstitution of H1-depleted fibroblast nuclei with nonphosphorylated or phosphorylated H5 was performed at physiological ionic strength. The bound H5 was then extracted using NaCl concentrations in the range of 0.15 to 0.7 M. The release of the H5 molecules was monitored by DAPI staining and image cytofluorometry. Our results show that H5 phosphorylation substantially reduced its affinity for chromatin in situ, which support previous observations indicating that C-terminal phosphorylation may be essential for the biological functions of linker histones. © 2012 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]
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- 2013
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108. Application of high-performance capillary electrophoresis to the analysis of H1 histones
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Lindner, Herbert, primary, Wurm, Martin, additional, Dirschlmayer, Arnold, additional, Sarg, Bettina, additional, and Helliger, Wilfried, additional
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- 1993
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109. Separation of phosphorylated histone H1 variants by high-performance capillary electrophoresis
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Lindner, Herbert, primary, Helliger, Wilfried, additional, Dirschlmayer, Arnold, additional, Talasz, Heribert, additional, Wurm, Martin, additional, Sarg, Bettina, additional, Jaquemar, Markus, additional, and Puschendorf, Bernd, additional
- Published
- 1992
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110. Preparation via coligand exchange and characterization of [99mTc-EDDA-HYNIC-D-Phe1,Tyr3]Octreotide (99mTc-EDDA/HYNIC-TOC).
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von Guggenberg, Elisabeth, Sarg, Bettina, Lindner, Herbert, Melendez Alafort, Laura, Mather, Stephen J., Moncayo, Roy, and Decristoforo, Clemens
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- 2003
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111. EspP, a serine protease of enterohaemorrhagic Escherichia coli, impairs complement activation by cleaving complement factors C3/C3b and C5
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Orth, Dorothea, Ehrlenbach, Silvia, Brockmeyer, Jens, Khan, Abdul Basit, Huber, Georg, Karch, Helge, Sarg, Bettina, Lindner, Herbert, and Würzner, Reinhard
- Published
- 2010
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112. Preparation via coligand exchange and characterization of [<SUP>99m</SUP>Tc-EDDA-HYNIC-D-Phe<SUP>1</SUP>,Tyr<SUP>3</SUP>]Octreotide (<SUP>99m</SUP>TcEDDA/HYNICTOC)
- Author
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Guggenberg, Elisabeth von, Sarg, Bettina, Lindner, Herbert, Alafort, Laura Melendez, Mather, Stephen J., Moncayo, Roy, and Decristoforo, Clemens
- Abstract
[99mTc-EDDAHYNIC-D-Phe1,Tyr3]octreotide (99mTc-EDDA/HYNICTOC) is a promising new agent with the potential to replace [111In-DTPA-D-Phe1]-octreotide in somatostatin receptor scintigraphy. This hydrazinonicotinic acid derivatized somatostatin complex contains ethylenediamine N,N' diacetic acid (EDDA) as a coligand resulting in a high in vitro and in vivo stability. Since direct 99mTc-labelling of HYNICTOC with EDDA results in low labelling yields, in this study we describe the preparation of 99mTc-EDDA/HYNIC-TOC via coligand exchange from Tricine for EDDA. Exchange of coligands is achieved at elevated temperature and under optimized conditions of pH, EDDA concentration and stannous ion. High labelling yields (mean 95.9%) were achieved at high specific activities (>37GBq/µmol). Characterization via HPLC, receptor binding and LCMS of the resulting complex is described. The formulation developed enables rapid and simple labelling of 99mTc-EDDA/HYNICTOC in a manner suitable for a clinical setting. Copyright © 2003 John Wiley & Sons, Ltd.
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- 2003
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113. 182 Complement C7 and clusterin form a stable complex in circulation.
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Massri, Mariam, Toonen, Erik J.M., Sarg, Bettina, Kremser, Leopold, Grasse, Marco, Skjoedt, Mikkel-Ole, Bayarri-Olmos, Raffael, Rosbjerg, Anne, Garred, Peter, Orth, Dorothea, Prohászka, Zoltán, and Würzner, Reinhard
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COMPLEMENT (Immunology) , *CLUSTERIN - Published
- 2023
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114. PROTEOMIC IDENTIFICATION OF NOVEL BIOMARKERS FOR DONOR SPECIFIC IMMUNE TOLERANCE.
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Kienzl, Katrin, Sarg, Bettina, Golderer, Georg, Lindner, Herbert, Werner, Ernst R, Werner-Felmayer, Gabriele, Maglione, Manuel, Schneeberger, Stefan, Margreiter, Raimund, and Brandacher, Gerald
- Published
- 2006
115. DETECTION OF BIOMARKERS FOR ACUTE CARDIAC ALLOGRAFT REJECTION BY PROTEOMIC ANALYSIS.
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Kienzl, Katrin, Sarg, Bettina, Golderer, Georg, Lindner, Herbert, Werner, Ernst R, Maglione, Manuel, Schneeberger, Stefan, Margreiter, Raimund, and Brandacher, Gerald
- Published
- 2006
116. A+-Helix of Protein C Inhibitor (PCI) Is a Cell-penetrating Peptide That Mediates Cell Membrane Permeation of PCI.
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Hanjiang Yang, Wahlmüller, Felix Christof, Sarg, Bettina, Furtmüller, Margareta, and Geiger, Margarethe
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PROTEIN C , *BLOOD coagulation factors , *CELL membranes , *PROTEOLYTIC enzymes , *SERPINS - Abstract
Protein C inhibitor (PCI) is a serpin with broad protease reactivity. It binds glycosaminoglycans and certain phospholipids that can modulate its inhibitory activity. PCI can penetrate through cellular membranes via binding to phosphatidylethanolamine. The exact mechanism of PCI internalization and the intracellular role of the serpin are not well understood. Here we showed that testisin, a glycosylphosphatidylinositol-anchored serine protease, cleaved human PCI and mouse PCI (mPCI) at their reactive sites as well as at sites close to their N terminus. This cleavage was observed not only with testisin in solution but also with cell membrane-anchored testisin on U937 cells. The cleavage close to the N terminus released peptides rich in basic amino acids. Synthetic peptides corresponding to the released peptides of human PCI (His¹-Arg11) and mPCI (Arg¹-Ala18) functioned as cell-penetrating peptides. Because intact mPCI but not testisin-cleaved mPCI was internalized by Jurkat T cells, a truncated mPCI mimicking testisin-cleaved mPCI was created. The truncated mPCI lacking 18 amino acids at the N terminus was not taken up by Jurkat T cells. Therefore our model suggests that testisin or other proteases could regulate the internalization of PCI by removing its N terminus. This may represent one of the mechanisms regulating the intracellular functions of PCI. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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117. Expression of transport proteins in the rete mirabile of european silver and yellow eel.
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Schneebauer, Gabriel, Drechsel, Victoria, Dirks, Ron, Faserl, Klaus, Sarg, Bettina, and Pelster, Bernd
- Abstract
Background: In physoclist fishes filling of the swimbladder requires acid secretion of gas gland cells to switch on the Root effect and subsequent countercurrent concentration of the initial gas partial pressure increase by back-diffusion of gas molecules in the rete mirabile. It is generally assumed that the rete mirabile functions as a passive exchanger, but a detailed analysis of lactate and water movements in the rete mirabile of the eel revealed that lactate is diffusing back in the rete. In the present study we therefore test the hypothesis that expression of transport proteins in rete capillaries allows for back-diffusion of ions and metabolites, which would support the countercurrent concentrating capacity of the rete mirabile. It is also assumed that in silver eels, the migratory stage of the eel, the expression of transport proteins would be enhanced. Results: Analysis of the transcriptome and of the proteome of rete mirabile tissue of the European eel revealed the expression of a large number of membrane ion and metabolite transport proteins, including monocarboxylate and glucose transport proteins. In addition, ion channel proteins, Ca2+-ATPase, Na+/K+-ATPase and also F1F0-ATP synthase were detected. In contrast to our expectation in silver eels the expression of these transport proteins was not elevated as compared to yellow eels. A remarkable number of enzymes degrading reactive oxygen species (ROS) was detected in rete capillaries. Conclusions: Our results reveal the expression of a large number of transport proteins in rete capillaries, so that the back diffusion of ions and metabolites, in particular lactate, may significantly enhance the countercurrent concentrating ability of the rete. Metabolic pathways allowing for aerobic generation of ATP supporting secondary active transport mechanisms are established. Rete tissue appears to be equipped with a high ROS defense capacity, preventing damage of the tissue due to the high oxygen partial pressures generated in the countercurrent system. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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118. Identification of voltage-gated K+ channel beta 2 (Kvβ2) subunit as a novel interaction partner of the pain transducer Transient Receptor Potential Vanilloid 1 channel (TRPV1).
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Bavassano, Carlo, Marvaldi, Letizia, Langeslag, Michiel, Sarg, Bettina, Lindner, Herbert, Klimaschewski, Lars, Kress, Michaela, Ferrer-Montiel, Antonio, and Knaus, Hans-Günther
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ION channel gating mechanisms , *POTASSIUM channels , *TRP channels , *INFLAMMATION , *PAIN perception , *GANGLIA - Abstract
Abstract: The Transient Receptor Potential Vanilloid 1 (TRPV1, vanilloid receptor 1) ion channel plays a key role in the perception of thermal and inflammatory pain, however, its molecular environment in dorsal root ganglia (DRG) is largely unexplored. Utilizing a panel of sequence-directed antibodies against TRPV1 protein and mouse DRG membranes, the channel complex from mouse DRG was detergent-solubilized, isolated by immunoprecipitation and subsequently analyzed by mass spectrometry. A number of potential TRPV1 interaction partners were identified, among them cytoskeletal proteins, signal transduction molecules, and established ion channel subunits. Based on stringent specificity criteria, the voltage-gated K+ channel beta 2 subunit (Kvβ2), an accessory subunit of voltage-gated K+ channels, was identified of being associated with native TRPV1 channels. Reverse co-immunoprecipitation and antibody co-staining experiments confirmed TRPV1/Kvβ2 association. Biotinylation assays in the presence of Kvβ2 demonstrated increased cell surface expression levels of TRPV1, while patch-clamp experiments resulted in a significant increase of TRPV1 sensitivity to capsaicin. Our work shows, for the first time, the association of a Kvβ subunit with TRPV1 channels, and suggests that such interaction may play a role in TRPV1 channel trafficking to the plasma membrane. [Copyright &y& Elsevier]
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- 2013
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119. ICn159 Folds into a Pleckstrin Homology Domain-like Structure.
- Author
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Fürst, Johannes, Schedlbauer, Andreas, Gandin, Rosaria, Garavaglian, Maria Lisa, Stefano Saino, Gschwentner, Martin, Sarg, Bettina, Lindned, Herbert, Jakab, Martin, Ritter, Markus, Bazzini, Claudia, Botta, Guido, Meyer, Giuliano, Kontaxis, Georg, Tilly, Ben C., Konrat, Robert, and Paulmichl, Markus
- Subjects
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RNA splicing , *GENETIC regulation , *HOMOLOGY (Biology) , *CELL membranes , *ION channels , *LIPIDS , *PROTEIN kinase C , *CYCLIC adenylic acid - Abstract
ICln is a multifunctional protein involved in regulatory mechanisms as different as membrane ion transport and RNA splicing. The protein is water-soluble, and during regulatory volume decrease after cell swelling, it is able to migrate from the cytosol to the cell membrane. Purified, water-soluble ICln is able to insert into lipid bilayers to form ion channels. Here, we show that ICln159, a truncated ICln mutant, which is also able to form ion channels in lipid bilayers, belongs to the pleckstrin homology (PH) domain superfold family of proteins. The ICln PH domain shows unusual properties as it lacks the electrostatic surface polarization seen in classical PH domains. However, similar to many classical PH domain-containing proteins, ICln interacts with protein kinase C, and in addition, interacts with cAMP-dependent protein kinase and cGMP-dependent protein kinase type II but not cGMP-dependent protein kinase type Iβ. A major phosphorylation site for all three kinases is Ser-45 within the ICln PH domain. Furthermore, ICln159 interacts with LSm4, a protein involved in splicing and mRNA degradation, suggesting that the ICln159 PH domain may serve as a protein-protein interaction platform. [ABSTRACT FROM AUTHOR]
- Published
- 2005
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120. Regulation and Processing of Maize Histone Deacetylase Hda1 by Limited Proteolysis.
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Pipal, Alexandra, Goralik-Schramel, Maria, Lusser, Alexandra, Lanzanova, Chiara, Sarg, Bettina, Loidl, Adele, Lindner, Herbert, Rossi, Vincenzo, and Loidl, Peter
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YEAST , *PROTEOLYSIS , *AMINO acid sequence , *PROTEIN analysis , *PEPTIDES - Abstract
Reports on the processing of the yeast Hda1-p84 to enzymatically active Hda1-p48 by limited proteolysis. Comparison of the amino acid sequences of ZmHda1 and related proteins; Expression of ZmHda1 mRNA in different maize organs and at various germination stages of embryos; Amino acid sequences determined by protein microsequencing of peptides from highly purified ZmHda1-p48.
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- 2003
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121. Truncated variants of MAGEL2 are involved in the etiologies of the Schaaf-Yang and Prader-Willi syndromes.
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Heimdörfer D, Vorleuter A, Eschlböck A, Spathopoulou A, Suarez-Cubero M, Farhan H, Reiterer V, Spanjaard M, Schaaf CP, Huber LA, Kremser L, Sarg B, Edenhofer F, Geley S, de Araujo MEG, and Huettenhofer A
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- Humans, Chromosomes, Human, Pair 15 genetics, Cytoplasm metabolism, HEK293 Cells, Mutation, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Proteins genetics, Proteins metabolism, RNA, Small Nucleolar genetics, Intracellular Signaling Peptides and Proteins, Intrinsically Disordered Proteins, Prader-Willi Syndrome genetics
- Abstract
The neurodevelopmental disorders Prader-Willi syndrome (PWS) and Schaaf-Yang syndrome (SYS) both arise from genomic alterations within human chromosome 15q11-q13. A deletion of the SNORD116 cluster, encoding small nucleolar RNAs, or frameshift mutations within MAGEL2 result in closely related phenotypes in individuals with PWS or SYS, respectively. By investigation of their subcellular localization, we observed that in contrast to a predominant cytoplasmic localization of wild-type (WT) MAGEL2, a truncated MAGEL2 mutant was evenly distributed between the cytoplasm and the nucleus. To elucidate regulatory pathways that may underlie both diseases, we identified protein interaction partners for WT or mutant MAGEL2, in particular the survival motor neuron protein (SMN), involved in spinal muscular atrophy, and the fragile-X-messenger ribonucleoprotein (FMRP), involved in autism spectrum disorders. The interactome of the non-coding RNA SNORD116 was also investigated by RNA-CoIP. We show that WT and truncated MAGEL2 were both involved in RNA metabolism, while regulation of transcription was mainly observed for WT MAGEL2. Hence, we investigated the influence of MAGEL2 mutations on the expression of genes from the PWS locus, including the SNORD116 cluster. Thereby, we provide evidence for MAGEL2 mutants decreasing the expression of SNORD116, SNORD115, and SNORD109A, as well as protein-coding genes MKRN3 and SNRPN, thus bridging the gap between PWS and SYS., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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122. The cytosolic form of dual localized BolA family protein Bol3 is important for adaptation to iron starvation in Aspergillus fumigatus .
- Author
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Oberegger S, Misslinger M, Faserl K, Sarg B, Farhan H, and Haas H
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- Adaptation, Physiological, Cell Nucleus metabolism, Protein Transport, Proteomics methods, Iron-Sulfur Proteins metabolism, Iron-Sulfur Proteins genetics, Gene Expression Regulation, Fungal, Acetylation, Aspergillus fumigatus metabolism, Aspergillus fumigatus genetics, Fungal Proteins metabolism, Fungal Proteins genetics, Cytosol metabolism, Mitochondria metabolism, Iron metabolism
- Abstract
Aspergillus fumigatus is the predominant mould pathogen for humans. Adaption to host-imposed iron limitation has previously been demonstrated to be essential for its virulence. [2Fe-2S] clusters are crucial as cofactors of several metabolic pathways and mediate cytosolic/nuclear iron sensing in fungi including A. fumigatus . [2Fe-2S] cluster trafficking has been shown to involve BolA family proteins in both mitochondria and the cytosol/nucleus. Interestingly, both A. fumigatus homologues, termed Bol1 and Bol3, possess mitochondrial targeting sequences, suggesting the lack of cytosolic/nuclear versions. Here, we show by the combination of mutational, proteomic and fluorescence microscopic analyses that expression of the Bol3 encoding gene leads to dual localization of gene products to mitochondria and the cytosol/nucleus via alternative translation initiation downstream of the mitochondrial targeting sequence, which appears to be highly conserved in various Aspergillus species. Lack of either mitochondrial Bol1 or Bol3 was phenotypically inconspicuous while lack of cytosolic/nuclear Bol3 impaired growth during iron limitation but not iron sensing which indicates a particular importance of [2Fe-2S] cluster trafficking during iron limitation. Remarkably, cytosolic/nuclear Bol3 differs from the mitochondrial version only by N-terminal acetylation, a finding that was only possible by mutational hypothesis testing.
- Published
- 2024
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123. Silicone implant surface microtopography modulates inflammation and tissue repair in capsular fibrosis.
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Schoberleitner I, Faserl K, Tripp CH, Pechriggl EJ, Sigl S, Brunner A, Zelger B, Hermann-Kleiter N, Baier L, Steinkellner T, Sarg B, Egle D, Brunner C, and Wolfram D
- Subjects
- Humans, Silicones, Fibrosis, Wound Healing, Prostheses and Implants, Inflammation
- Abstract
Excessive fibrous capsule formation around silicone mammary implants (SMI) involves immune reactions to silicone. Capsular fibrosis, a common SMI complication linked to host responses, worsens with specific implant topographies. Our study with 10 patients investigated intra- and inter-individually, reduced surface roughness effects on disease progression, wound responses, chronic inflammation, and capsular composition. The results illuminate the significant impact of surface roughness on acute inflammatory responses, fibrinogen accumulation, and the subsequent fibrotic cascade. The reduction of surface roughness to an average roughness of 4 μm emerges as a promising approach for mitigating detrimental immune reactions, promoting healthy wound healing, and curbing excessive fibrosis. The identified proteins adhering to rougher surfaces shed light on potential mediators of pro-inflammatory and pro-fibrotic processes, further emphasizing the need for meticulous consideration of surface design. The composition of the implant capsule and the discovery of intracapsular HSP60 expression highlight the intricate web of stress responses and immune activation that can impact long-term tissue outcomes., 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 © 2024 Schoberleitner, Faserl, Tripp, Pechriggl, Sigl, Brunner, Zelger, Hermann-Kleiter, Baier, Steinkellner, Sarg, Egle, Brunner and Wolfram.)
- Published
- 2024
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124. Surface Topography, Microbial Adhesion, and Immune Responses in Silicone Mammary Implant-Associated Capsular Fibrosis.
- Author
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Schoberleitner I, Baier L, Lackner M, Zenz LM, Coraça-Huber DC, Ullmer W, Damerum A, Faserl K, Sigl S, Steinkellner T, Winkelmann S, Sarg B, Egle D, Brunner C, and Wolfram D
- Subjects
- Humans, Female, Silicones, Proteome, RNA, Ribosomal, 16S genetics, Mastectomy, Fibrosis, Breast Implants adverse effects, Breast Neoplasms surgery, Anti-Infective Agents
- Abstract
Breast cancer is the most common cancer in women globally, often necessitating mastectomy and subsequent breast reconstruction. Silicone mammary implants (SMIs) play a pivotal role in breast reconstruction, yet their interaction with the host immune system and microbiome remains poorly understood. This study investigates the impact of SMI surface topography on host antimicrobial responses, wound proteome dynamics, and microbial colonization. Biological samples were collected from ten human patients undergoing breast reconstruction with SMIs. Mass spectrometry profiles were analyzed for acute and chronic wound proteomes, revealing a nuanced interplay between topography and antimicrobial response proteins. 16S rRNA sequencing assessed microbiome dynamics, unveiling topography-specific variations in microbial composition. Surface topography alterations influenced wound proteome composition. Microbiome analysis revealed heightened diversity around rougher SMIs, emphasizing topography-dependent microbial invasion. In vitro experiments confirmed staphylococcal adhesion, growth, and biofilm formation on SMI surfaces, with increased texture correlating positively with bacterial colonization. This comprehensive investigation highlights the intricate interplay between SMI topography, wound proteome dynamics, and microbial transmission. The findings contribute to understanding host-microbe interactions on SMI surfaces, essential for optimizing clinical applications and minimizing complications in breast reconstruction.
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- 2024
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125. Complement C7 and clusterin form a complex in circulation.
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Massri M, Toonen EJM, Sarg B, Kremser L, Grasse M, Fleischer V, Torres-Quesada O, Hengst L, Skjoedt MO, Bayarri-Olmos R, Rosbjerg A, Garred P, Orth-Höller D, Prohászka Z, and Würzner R
- Subjects
- Complement System Proteins metabolism, Complement Membrane Attack Complex metabolism, Complement Activation, Complement C7 metabolism, Clusterin
- Abstract
Introduction: The complement system is part of innate immunity and is comprised of an intricate network of proteins that are vital for host defense and host homeostasis. A distinct mechanism by which complement defends against invading pathogens is through the membrane attack complex (MAC), a lytic structure that forms on target surfaces. The MAC is made up of several complement components, and one indispensable component of the MAC is C7. The role of C7 in MAC assembly is well documented, however, inherent characteristics of C7 are yet to be investigated., Methods: To shed light on the molecular characteristics of C7, we examined the properties of serum-purified C7 acquired using polyclonal and novel monoclonal antibodies. The properties of serum‑purified C7 were investigated through a series of proteolytic analyses, encompassing Western blot and mass spectrometry. The nature of C7 protein-protein interactions were further examined by a novel enzyme-linked immunosorbent assay (ELISA), as well as size‑exclusion chromatography., Results: Protein analyses showcased an association between C7 and clusterin, an inhibitory complement regulator. The distinct association between C7 and clusterin was also demonstrated in serum-purified clusterin. Further assessment revealed that a complex between C7 and clusterin (C7-CLU) was detected. The C7-CLU complex was also identified in healthy serum and plasma donors, highlighting the presence of the complex in circulation., Discussion: Clusterin is known to dissociate the MAC structure by binding to polymerized C9, nevertheless, here we show clusterin binding to the native form of a terminal complement protein in vivo. The presented data reveal that C7 exhibits characteristics beyond that of MAC assembly, instigating further investigation of the effector role that the C7-CLU complex plays in the complement cascade., Competing Interests: ET is an employee of Hycult Biotech. The remaining 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. The reviewer AZ declared a shared affiliation with the author M-OS to the handling editor at the time of review., (Copyright © 2024 Massri, Toonen, Sarg, Kremser, Grasse, Fleischer, Torres-Quesada, Hengst, Skjoedt, Bayarri-Olmos, Rosbjerg, Garred, Orth-Höller, Prohászka and Würzner.)
- Published
- 2024
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126. Skeletal muscle proteome analysis underpins multifaceted mitochondrial dysfunction in Friedreich's ataxia.
- Author
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Indelicato E, Faserl K, Amprosi M, Nachbauer W, Schneider R, Wanschitz J, Sarg B, and Boesch S
- Abstract
Friedreich's ataxia (FRDA) is a severe multisystemic disorder caused by a deficiency of the mitochondrial protein frataxin. While some aspects of FRDA pathology are developmental, the causes underlying the steady progression are unclear. The inaccessibility of key affected tissues to sampling is a main hurdle. Skeletal muscle displays a disease phenotype and may be sampled in vivo to address open questions on FRDA pathophysiology. Thus, we performed a quantitative mass spectrometry-based proteomics analysis in gastrocnemius skeletal muscle biopsies from genetically confirmed FRDA patients ( n = 5) and controls. Obtained data files were processed using Proteome Discoverer and searched by Sequest HT engine against a UniProt human reference proteome database. Comparing skeletal muscle proteomics profiles between FRDA and controls, we identified 228 significant differentially expressed (DE) proteins, of which 227 were downregulated in FRDA. Principal component analysis showed a clear separation between FRDA and control samples. Interactome analysis revealed clustering of DE proteins in oxidative phosphorylation, ribosomal elements, mitochondrial architecture control, and fission/fusion pathways. DE findings in the muscle-specific proteomics suggested a shift toward fast-twitching glycolytic fibers. Notably, most DE proteins (169/228, 74%) are target of the transcription factor nuclear factor-erythroid 2. Our data corroborate a mitochondrial biosignature of FRDA, which extends beyond a mere oxidative phosphorylation failure. Skeletal muscle proteomics highlighted a derangement of mitochondrial architecture and maintenance pathways and a likely adaptive metabolic shift of contractile proteins. The present findings are relevant for the design of future therapeutic strategies and highlight the value of skeletal muscle-omics as disease state readout in FRDA., 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 Indelicato, Faserl, Amprosi, Nachbauer, Schneider, Wanschitz, Sarg and Boesch.)
- Published
- 2023
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127. Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers.
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Kellnerová S, Huber S, Massri M, Fleischer V, Losso K, Sarg B, Kremser L, Talasz H, He X, Varrone E, Brigotti M, Ardissino G, Orth-Höller D, and Würzner R
- Abstract
Shiga toxins (Stxs), especially the Stx2a subtype, are the major virulence factors involved in enterohemorrhagic Escherichia coli (EHEC)-associated hemolytic uremic syndrome (eHUS), a life-threatening disease causing acute kidney injury, especially in children. After oral transmission and colonization in the gut, EHEC release Stx. Intracellular cleavage of the Stx A subunit, when followed by reduction, boosts the enzymatic activity that causes damage to targeted cells. This cleavage was assumed to be mostly mediated by furin during Stx intracellular trafficking. To investigate whether this cleavage could occur in the intestine, even prior to entering target cells, Stx2a A subunit structure (intact or cleaved) was characterized after its exposure to specific host factors present in human stool. The molecular weight of Stx2a A subunit/fragments was determined by immunoblotting after electrophoretic separation under reducing conditions. In this study, it was demonstrated that Stx2a is cleaved by certain human stool components. Trypsin and chymotrypsin-like elastase 3B (CELA3B), two serine proteases, were identified as potential candidates that can trigger the extracellular cleavage of Stx2a A subunit directly after its secretion by EHEC in the gut. Whether the observed cleavage indeed translates to natural infections and plays a role in eHUS pathogenesis has yet to be determined. If so, it seems likely that a host's protease profile could affect disease development by changing the toxin's biological features.
- Published
- 2023
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128. Protein sites of attack of N-chlorotaurine in Escherichia coli.
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Arnitz R, Sarg B, Ott HW, Neher A, Lindner H, and Nagl M
- Subjects
- Acyl-Carrier Protein S-Malonyltransferase metabolism, Bacterial Proteins drug effects, Chaperonin 60 metabolism, Cysteine chemistry, Electrophoresis, Gel, Two-Dimensional, Oxidation-Reduction, Periplasmic Binding Proteins metabolism, Ribosomal Proteins metabolism, Spectrometry, Mass, Electrospray Ionization, Sulfonic Acids metabolism, Taurine pharmacology, Enzyme Inhibitors pharmacology, Escherichia coli drug effects, Escherichia coli metabolism, Protein Serine-Threonine Kinases antagonists & inhibitors, Proteomics, Taurine analogs & derivatives
- Abstract
N-Chlorotaurine sodium (NCT) is a promising microbicidal agent for topical treatment of infections. Its targets of attack in Escherichia coli have been investigated by proteomics. Incubation in 1% NCT for 10 and 30 min revealed a change of the charge and a separation of numerous proteins into a series of spots with a different pI. Charge differences could be related to oxidation of cysteine residues to their corresponding sulfonic acids. Heat shock protein 60 appeared, while ribosome-releasing factor, d-ribose periplasmic binding protein, and malonyl-CoA transacylase spots decreased. These results indicate penetration of oxidation capacity into the bacteria and destruction of essential proteins by NCT.
- Published
- 2006
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129. Calgranulins in cystic fluid and serum from patients with ovarian carcinomas.
- Author
-
Ott HW, Lindner H, Sarg B, Mueller-Holzner E, Abendstein B, Bergant A, Fessler S, Schwaerzler P, Zeimet A, Marth C, and Illmensee K
- Subjects
- Adult, Aged, Aged, 80 and over, Calgranulin A blood, Calgranulin B blood, Female, Humans, Middle Aged, Ovarian Cysts blood, Ovarian Cysts metabolism, Ovarian Neoplasms blood, Calgranulin A metabolism, Calgranulin B metabolism, Cyst Fluid metabolism, Ovarian Neoplasms metabolism
- Abstract
Ovarian cancer remains still associated with poor prognosis because it is diagnosed predominantly at advanced stages. Ovarian-specific tumor markers do not yet exist for early detection of the disease. At the search of diagnostic markers for ovarian cancer, proteomic-based approaches have focused on novel investigations of neoplastic processes in tumor patients. Cystic fluids of malignant and benign ovarian tumors and serum from the corresponding patients were collected and processed for two-dimensional gel electrophoresis. Proteins were visualized on the gels by silver staining. At the low molecular mass level between 10 and 20 kDa, selected protein spots were additionally processed for nanospray mass spectrometry and partial amino acid sequencing. For protein identification, the sequencing results were compared with computer information from a protein data bank. Protein patterns from cystic fluids of ovarian carcinomas differed significantly from those of benign cysts and revealed additional polypeptides at low molecular mass level between 10 and 20 kDa. Protein patterns from serum of patients with malignant ovarian tumors also contained additional polypeptides between 10 and 20 kDa that were not detected in serum from patients with benign cysts. The additional proteins in serum were present in similar electrophoretic positions compared with those found in the cystic fluid of the corresponding ovarian carcinomas. Protein spots in the range of 10-20 kDa were selected for partial amino acid sequencing. Two protein spots were identified as calgranulin A and three spots as calgranulin B. Either both proteins or only calgranulin A or B were present in cystic fluid from ovarian carcinomas and serum of the corresponding patients. These two proteins were absent or not detectable in fluid from benign ovarian cysts and in serum from those patients. Our investigations concerning protein patterns in cystic fluid of malignant and benign ovarian tumors provide new information about alterations in protein synthesis linked to neoplastic events of the ovary. With the proteomic strategy, new tumor markers are characterized and may serve for diagnostic purposes of patients with ovarian cancer.
- Published
- 2003
130. Capillary electrophoresis analysis of histones, histone variants, and their post-translationally modified forms: a review.
- Author
-
Lindner H, Sarg B, and Helliger W
- Subjects
- Buffers, Histones chemistry, Hydrogen-Ion Concentration, Electrophoresis, Capillary methods, Histones analysis, Histones metabolism, Protein Processing, Post-Translational physiology
- Abstract
Histones are the most intensively studied group of basic nuclear proteins and are of great importance with regard to the organization of chromatin structure and control of gene activity. The complexity of this protein family places very high demands on the analytical methods applied. The advent of high-performance capillary electrophoresis provided a promising new tool for their separation, which is an essential prerequisite for studying the biological role of this protein family. Problems inherent to histone analysis due to their unique physical and chemical properties are reviewed, and the pros and cons of distinct strategies developed for CE separation of these proteins are discussed.
- Published
- 2003
131. Anti-SLA seropositive autoimmune hepatitis sera recognize distinct subunits of glutathione S-transferase: high prevalence of the Ya autoantigen.
- Author
-
Wesierska-Gadek J, Lindner H, Hitchman E, Sarg B, and Penner E
- Subjects
- Animals, Chromatography, Affinity, Chromatography, High Pressure Liquid, Electrophoresis, Polyacrylamide Gel, Hepatitis, Autoimmune enzymology, Humans, Isoenzymes immunology, Rats, Autoantibodies, Autoantigens immunology, Glutathione Transferase immunology, Hepatitis, Autoimmune immunology
- Abstract
We have recently reported that anti-SLA seropositive autoimmune hepatitis (AIH) patients develop autoantibodies against glutathione S-transferase (GST). GSTs are multifunctional enzymes mediating hepatic detoxification of cytotoxic and genotoxic compounds and are also involved in biliary secretion. We have observed varying reactivity of individual AIH sera towards several GST isoenzymes. Since the GST subunits have very similar molar masses and therefore are not satisfactorily resolved by one-dimensional gel electrophoresis, we have performed their fractionation by reverse-phase high performance liquid chromatography (HPLC) to better separate the individual GST isoenzymes. 4 individual GST subunits were isolated as judged by electrophoretic analysis of the 4 distinct peaks. The identity of isolated proteins was unequivocally determined by protein sequencing. Isolated subtypes were loaded on 15% SDS gels and blotted. Immunoblotting was performed with eleven anti-SLA positive sera that displayed differential reactivity with total GSTs. Fractionation of the GSTs by HPLC did not impair their ability to react with specific autoantibodies. Interestingly, the majority of GST-positive AIH sera reacted with one or two GST subtypes, only two sera recognized 3 subunits. Ya was most prevalent autoantigen. Autoantibodies against Yb2 were detected solely in one serum. This pattern of reactivity indicates that individual patients' sera discriminate between GST subunits despite their sequence homology. It is well known that the GST variants differ within their amino-terminal part while the residual moiety is highly conserved. It would suggest that autoantibodies recognize distinct epitopes located within amino-terminus of individual GST variants.
- Published
- 2002
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