Comparative studies of Ca2+-dependent proteases (CDP I and CDP II) from Allomyces arbuscula

Article Article scientifique Article Comparative studies of Ca2+-dependent proteases (CDP I and CDP II) from Allomyces arbuscula eng 1999-07 author Ojha Mukti 25001 author Cattaneo Arlette 26052 author Schwendimann Bernard 25349 765-770 10.1016/S0300-9084(99)80135-7 0300-9084 https://linkinghub.elsevier.com/retrieve/pii/S0300908499801357 Allomyces arbuscula, an aquatic fungus, contains two Ca2+-dependent neutral cysteine proteases (CDP I and CDP II), eluting respectively, at 0.07 and 0.2 M NaCl from DEAE cellulose columns. The purified CDP I has a Mr of 39 kDa whereas CDP II appears as a doublet of 43 and 40 kDa. Both enzymes require free thiol, the same concentration of Ca2+ for half maximal activation, and are inactivated by thiol protease inhibitors. Our results show that despite these similarities the two enzymes are different because affinity-purified CDP II antibodies do not cross-react with CDP I antigen in Western blots. In contrast, there is a strong cross-reaction between the two 43 and 40 kDa CDP II peptides and their respective antibodies. Both enzymes cleave preferentially the carboxy terminus of Arg and to a limited extent Lys on the cleavage site. This primary specificity is governed by the nature of the amino acids in the P2 and P3 positions. In general either Pro or Gly in P2 is required, with preference for Pro and in P3 position, Gly over Val. CDP II has higher catalytic activity than CDP I. The sulfhydryl reagent NEM is a more potent inhibitor of CDP I than CDP II. Although the function of the phosphorylable site(s) is not clear, both CDP I and CDP II contain phosphorylable serine residue(s). Dewey 580 45 Département des sciences végétales 163 false Biochimie 81 7 b5b80e11-c6bc-4c21-b0c8-79c1f80fa0c9 Article (Published version) Restricted 1-s2.0-S0300908499801357-main.pdf 149551 application/pdf 55cbf1dc5cb7dc0e03ae179d2c55c0ab35054b70c6681e0c6a178c520b471531 restrictedAccess Faculté des sciences / Section de biologie / Département des sciences végétales

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Comparative studies of Ca²⁺-dependent proteases (CDP I and CDP II) from Allomyces arbuscula	Biochimie			1999	20	0

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Schwendimann, Bernard