Regulation of the V-type ATPase by redox modulation
| Autor: | Thorsten Seidel, Melanie Krebs, Stefan Scholl, Karin Schumacher, Karl-Josef Dietz, Irene Marten, Rainer Hedrich, Florian Rienmüller, Patricia Janetzki, Miriam Hanitzsch |
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| Rok vydání: | 2012 |
| Předmět: |
Models
Molecular Vacuolar Proton-Translocating ATPases Patch-Clamp Techniques Protein Conformation ATPase Protein subunit Molecular Sequence Data Arabidopsis Oxidative phosphorylation Biochemistry Dithiothreitol chemistry.chemical_compound V-ATPase Amino Acid Sequence Cysteine Molecular Biology health care economics and organizations Conserved Sequence chemistry.chemical_classification biology Sequence Homology Amino Acid Chemistry Arabidopsis Proteins Cell Biology Hydrogen-Ion Concentration biology.organism_classification Plants Genetically Modified Recombinant Proteins Amino acid Protein Subunits Amino Acid Substitution biology.protein Mutagenesis Site-Directed Oxidation-Reduction |
| Zdroj: | The Biochemical journal. 448(2) |
| ISSN: | 1470-8728 |
| Popis: | ATP-hydrolysis and proton pumping by the V-ATPase (vacuolar proton-translocating ATPase) are subject to redox regulation in mammals, yeast and plants. Oxidative inhibition of the V-ATPase is ascribed to disulfide-bond formation between conserved cysteine residues at the catalytic site of subunit A. Subunits containing amino acid substitutions of one of three conserved cysteine residues of VHA-A were expressed in a vha-A null mutant background in Arabidopsis . In vitro activity measurements revealed a complete absence of oxidative inhibition in the transgenic line expressing VHA-A C256S, confirming that Cys256 is necessary for redox regulation. In contrast, oxidative inhibition was unaffected in plants expressing VHA-A C279S and VHA-A C535S, indicating that disulfide bridges involving these cysteine residues are not essential for oxidative inhibition. In vivo data suggest that oxidative inhibition might not represent a general regulatory mechanism in plants. Abbreviations: AM, acetoxymethyl ester; BCECF, 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein; BTP, bis-Tris propane; DTT, dithiothreitol; GSNO, nitrosoglutathione; p-PDM, N,N′-p-phenylbismaleimide; V-ATPase, vacuolar proton-translocating ATPase |
| Databáze: | OpenAIRE |
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