CYSTEINES OF CHLOROPLAST NADP-MALATE DEHYDROGENASE FORM MIXED DISULFIDES

Autor(en): OCHERETINA, O
SCHEIBE, R 
Stichwörter: Biochemistry & Molecular Biology; Biophysics; Cell Biology; CLONING; DEPENDENT REGULATORY SITE; DIRECTED MUTAGENESIS; EXPRESSION; GLUTATHIONE; LIMITED PROTEOLYSIS; MIXED DISULFIDE; NADP-MALATE DEHYDROGENASE; OXIDATIVE MODIFICATION; OXIDATIVE STRESS; PEA; PEA CHLOROPLAST; PURIFICATION; REDOX MODIFICATION; REDUCTIVE ACTIVATION; SEQUENCE; STAPHYLOCOCCUS AUREUS PROTEASE V8
Erscheinungsdatum: 1994
Herausgeber: WILEY
Journal: FEBS LETTERS
Volumen: 355
Ausgabe: 3
Startseite: 254
Seitenende: 258
Zusammenfassung: 
Chloroplast NADP-malate dehydrogenase (NADP-MDH) from pea and from spinach was N-terminally truncated by limited proteolysis with Staphylococcus aureus protease V8. The resulting monomeric enzymes lacking, respectively, the 37 and 38 N-terminal amino acids were inactive. Reduction and addition of low concentrations of guanidine-HCl (50-100 mM) resulted in a highly active enzyme of 850 units per mg protein. Equilibration of the truncated enzyme with various glutathione (GSH) redox buffers and assaying its activity in the presence of guanidine-HCl was used to establish the existence of protein-GSH mixed disulfides. This finding was further confirmed using incorporation of radioactively labelled thiol. The possible function of such cysteine modifications under oxidative stress and their regeneration by the thioredoxin system in the light is discussed.
ISSN: 18733468
DOI: 10.1016/0014-5793(94)01214-8

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