The influence of residue 190 in the S1 site of trypsin‐like serine proteases on substrate selectivity is universally conserved

FEBS Letters

Kopetzki, Erhard¹  Huber, Robert³  Hopfner, Karl-Peter²  Bode, Wolfram³  Brandstetter, Hans³  Sichler, Katrin³ 

[1] Roche Diagnostics GmbH, D-82372 Penzberg, Germany;Ludwig-Maximilians-Universität, Genzentrum, D-81377 Munich, Germany;Max-Planck-Institut für Biochemie, D-82152 Martinsried, Germany

关键词: Activity modulation;    Activity profile;    Coagulation enzyme;    Induced fit;    Hybrid protease;    Site-directed mutagenesis;    rf9a;    recombinant factor IXa;    rf10a;    recombinant factor Xa;    rTry;    recombinant trypsin;    rXYa;    recombinant factor Xa-trypsin hybrid;    pNA;    para-nitroanilide;    tGPR;    Chr TH;    tGPK;    Chr PL;    tGPX;    substrate pair tGPR and tGPK;    VLR;    S-2266;    VLK;    S-2251;    VLX;    substrate pair VLR and VLK;   

DOI  :  10.1016/S0014-5793(02)03495-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We examined the influence of Ser/Ala190 in the S1 site on P1 substrate selectivity in several serine proteases. The impact of residue 190 on the selectivity was constant, regardless of differences in original selectivity or reactivity. Substrate binding in S1 was optimised in all wild-type enzymes, while the effects on k _cat depended on the combination of residue 190 and substrate. Mutagenesis of residue 190 did not affect the S2–S4 sites. Pronounced selectivity for arginine residues was coupled with low enzymatic activity, in particular in recombinant factor IXa. This is due to the dominance of the S1–P1 interaction over substrate binding in the S2–S4 sites.

【 授权许可】

Unknown