Nardilysin cleaves peptides at monobasic sites

被引:17
作者
Chow, KM
Oakley, O
Goodman, J
Ma, ZL
Juliano, MA
Juliano, L
Hersh, LB
机构
[1] Univ Kentucky, Coll Med, Dept Mol & Cellular Biol, Lexington, KY 40536 USA
[2] Univ Kentucky, Coll Med, Lexington, KY 40536 USA
[3] Escola Paulista Med, Dept Biophys, Sao Paulo, Brazil
关键词
D O I
10.1021/bi027178d
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
Nardilysin (N-arginine dibasic convertase, EC 3.4.24.61) was first identified on the basis of its ability to cleave peptides containing an arginine dibasic pair, i.e., Arg-Arg or Arg-Lys. However, it was observed that an aromatic residue adjacent to the dibasic pair (i.e., Phe-Arg-Lys) could alter the cleavage site. In this study we determined whether nardilysin can cleave peptides at a single basic residue. Nardilysin cleaves beta-endorphin at the monobasic site, Phe(17)-Lys(18), with a k(cat)/K-m of 2 x 10(8) M-1 min(-1). This can be compared to a k(cat)/K-m of 8.5 x 10(8) M-1 min(-1) for cleavage between a dibasic pair in dynotphin B-13. Nardilysin also cleaves calcitonin at His-Arg and somatostatin-14 at Cys-Lys. We examined the hydrolysis of fluorogenic peptides based on the beta-endorphin 12-24 sequence, Abz-T-P-L-V-T-L-X1-X2N-A-I-I-K-Q-EDDnp. Nardilysin hydrolyzes the peptides when X-1-X-2 = F-K, F-R, W-K, M-K, Y-K, and L-K. The kinetics of cleavage at F-K and F-R are similar; however, K-F is not hydrolyzed. Nardilysin cleaves at two monobasic sites M-K and F-R of the kallidin model peptide AbzMISLMKRPPGFSPFRSSRI-NH2, releasing desArg(10) kallidin (KRPPGFSPF). However, nardilysin does not release desArg(10) kallidin from the physiological precursor low molecular weight kininogen. These studies extend the range of potential substrates for nardilysin and further substantiate that nardilysin is a true peptidase.
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页码:2239 / 2244
页数:6
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