2. Academic Validation
  3. Carboxypeptidase Y activity and maintenance is modulated by a large helical structure

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

  • FEBS Open Bio. 2019 Jul;9(7):1337-1343. doi: 10.1002/2211-5463.12686.
Mai Makino 1,  Takehiko Sahara 2,  Naoki Morita 3,  Hiroshi Ueno 4
Affiliations

Affiliations

  • 1 Department of Biochemistry, Nara Medical University, Kashihara, Japan.
  • 2 Bio-Design Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
  • 3 Molecular and Biological Technology Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo, Japan.
  • 4 Laboratory of Biochemistry and Applied Microbiology, School of Agriculture, Ryukoku University, Otsu, Japan.
PMID: 31173671 DOI: 10.1002/2211-5463.12686
Abstract

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.

Keywords

disulfide bond; protein folding; serine carboxypeptidase; α/β hydrolase fold.

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