Competitive inhibition analysis of the enzyme-substrate interaction in the carboxy-terminal processing of the precursor D1 protein of photosystem II reaction center using substituted oligopeptides.
A clear parallelism was demonstrated between the efficiency as substrate of the substituted oligopeptides corresponding to the carboxy-terminal (C-terminal) sequence of the precursor D1 protein (pD1) in the in vitro enzymatic assay and their competitive inhibitory capacity toward the proteolytic C-terminal processing of the full-length pD1 integrated in the intact photosystem II complex embedded in the thylakoid membrane of Scenedesmus obliquus LF-1 mutant, as shown e.g. by the influence of L343A, A345G and A345V substitutions and the effect of C-terminal fragments. This suggests that the basic mechanism for substrate recognition by the processing protease elucidated in the enzymatic analysis using synthetic oligopeptides is also effective in vivo, although it can sometimes be difficult to detect the consequence of amino acid substitution in the integrated systems.
Department of Biology, Okayama University, Japan.
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Y
Yamamoto
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K
Satoh
D020418:Algal Proteins; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001667:Binding, Competitive; D002268:Carboxypeptidases; D000460:Chlorophyta / Q000235:genetics; D002736:Chloroplasts / Q000201:enzymology / Q000235:genetics; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000494:pharmacology*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D043484:Proprotein Convertases; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D018724:Spinacia oleracea / Q000201:enzymology; D013379:Substrate Specificity
D020418:Algal Proteins; D009842:Oligopeptides; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D011498:Protein Precursors; D002268:Carboxypeptidases; D010450:Endopeptidases; D043484:Proprotein Convertases; C073491:photosystem II, D1 polypeptide carboxy-terminal processing protease