TY - JOUR
T1 - Nardilysin cleaves peptides at monobasic sites
AU - Chow, K. Martin
AU - Oakley, Oliver
AU - Goodman, Jack
AU - Ma, Zhangliang
AU - Aparecida Juliano, Maria
AU - Juliano, Luiz
AU - Hersh, Louis B.
PY - 2003/2/25
Y1 - 2003/2/25
N2 - 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 β-endorphin at the monobasic site, Phe17-Lys18, with a kcat/Km of 2 × 108 M-1 min-1. This can be compared to a kcat/Km of 8.5 × 108 M-1 min-1 for cleavage between a dibasic pair in dynorphin 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 β-endorphin 12-24 sequence, Abz-T-P-L-V-T-L-X1-X2-N-A-I-I-K-Q-EDDnp. Nardilysin hydrolyzes the peptides when X1-X2 = 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 Abz-MISLMKRPPGFSPFRSSRI-NH2, releasing desArg10 kallidin (KRPPGFSPF). However, nardilysin does not release desArg10 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.
AB - 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 β-endorphin at the monobasic site, Phe17-Lys18, with a kcat/Km of 2 × 108 M-1 min-1. This can be compared to a kcat/Km of 8.5 × 108 M-1 min-1 for cleavage between a dibasic pair in dynorphin 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 β-endorphin 12-24 sequence, Abz-T-P-L-V-T-L-X1-X2-N-A-I-I-K-Q-EDDnp. Nardilysin hydrolyzes the peptides when X1-X2 = 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 Abz-MISLMKRPPGFSPFRSSRI-NH2, releasing desArg10 kallidin (KRPPGFSPF). However, nardilysin does not release desArg10 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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U2 - 10.1021/bi027178d
DO - 10.1021/bi027178d
M3 - Article
C2 - 12590613
AN - SCOPUS:0037465338
SN - 0006-2960
VL - 42
SP - 2239
EP - 2244
JO - Biochemistry
JF - Biochemistry
IS - 7
ER -