Human kallikrein 6 activity is regulated via an autoproteolytic mechanism of activation/inactivation

被引:60
作者
Bayés, A
Tsetsenis, T
Ventura, S
Vendrell, J [1 ]
Aviles, FX
Sotiropoulou, G
机构
[1] Univ Autonoma Barcelona, Dept Bioquim & Biol Mol, E-08193 Barcelona, Spain
[2] Univ Autonoma Barcelona, Inst Biotecnol & Biomed, E-08193 Barcelona, Spain
[3] Univ Patras, Sch Hlth Sci, Dept Pharm, GR-26500 Patras, Greece
关键词
angiostatin; autoactivation; human kallikrein 6; serine protease; site-directed mutagenesis;
D O I
10.1515/BC.2004.061
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 (pro-hK6) 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.
引用
收藏
页码:517 / 524
页数:8
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