Carboxypeptidase Y activity and maintenance is modulated by a large helical structure

FEBS Open Bio

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[1] Bio‐Design Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan;Department of Biochemistry, Nara Medical University, Kashihara, Japan;Laboratory of Biochemistry and Applied Microbiology, School of Agriculture, Ryukoku University, Otsu, Japan;Molecular and Biological Technology Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo, Japan;

关键词: disulfide bond;    protein folding;    serine carboxypeptidase;    α/β hydrolase fold;   

DOI  :  10.1002/2211-5463.12686

来源: publisher

【 摘 要 】

Yeast carboxypeptidase Y (CPY) is a serine protease with broad substrate specificity. Structurally, CPY belongs to the α/β hydrolase fold family and contains characteristic large helices, termed the V‐shape helix, above the active site cavity. Four intramolecular disulfide bonds are located in and around the V‐shape helix. In this study, mutant CPYs were constructed in which one of these disulfide bonds was disrupted. Mutants lacking the C193–C207 bond located at the beginning of the V‐shape helix aggregated easily, while mutants lacking the C262–C268 bond located at the end of the V‐shape helix displayed decreased hydrolytic activity. The results indicate that the V‐shape helix is involved in CPY catalysis and in maintenance of its conformation.

【 授权许可】

Unknown