A. Bayés, T. Tsetsenis, S. Ventura, J. Vendrell, F.X. Aviles, G. Sotiropoulou
Human kallikrein 6 activity is regulated via an autoproteolytic mechanism of activation/inactivation
Human kallikrein 6 (protease M/zyme/neurosin) is a serine protease that has been suggested to be a serum biomarker for ovarian cancer and may also be involved in pathologies of the CNS. The precursor form of human kallikrein 6 (proh-K6) was overexpressed in Pichia pastoris and found to be autoprocessed to an active but unstable mature enzyme that subsequently yielded the inactive, self-cleavage product, hK6 (D[81]K[244]). Site-directed mutagenesis was used to investigate the basis for the intrinsic catalytic activity and the activation mechanism of pro-hK6. A single substitution R[80]->Q stabilized the activity of the mature enzyme, while substitution of the active site serine (S[197]->A) resulted in complete loss of hK6 proteolytic activity and facilitated protein production. Our data suggest that the enzymatic activity of hK6 is regulated by an autoactivation/autoinactivation mechanism. Mature hK6 displayed a trypsin-like activity against synthetic substrates and human plasminogen was identified as a putative physiological substrate for hK6, as specific cleavage at the plasminogen internal bond S[460]-V[461] resulted in the generation of angiostatin, an endogenous inhibitor of angiogenesis and metastatic growth.
Biological Chemistry, Walter de Gruyter
Print ISSN: 1431-6730
Volume: 385, 06/2004
Pages: 517 - 524
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