CDNA CLONING OF PORCINE BRAIN PROLYL ENDOPEPTIDASE AND IDENTIFICATION OF THE ACTIVE-SITE SERYL RESIDUE

被引:161
作者
RENNEX, D [1 ]
HEMMINGS, BA [1 ]
HOFSTEENGE, J [1 ]
STONE, SR [1 ]
机构
[1] FRIEDRICH MIESCHER INST,POSTFACH 2543,CH-4002 BASEL,SWITZERLAND
关键词
D O I
10.1021/bi00222a025
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
Prolyl endopeptidase is a cytoplasmic serine protease. The enzyme was purified from porcine kidney, and oligonucleotides based on peptide sequences from this protein were used to isolate a cDNA clone from a porcine brain library. This clone contained the complete coding sequence of prolyl endopeptidase and encoded a polypeptide with a molecular mass of 80751 Da. The deduced amino acid sequence of prolyl endopeptidase showed no sequence homology with other known serine proteases. [H-3]Diisopropyl fluorophosphate was used to identify the active-site serine of prolyl endopeptidase. One labeled peptide was isolated and sequenced. The sequence surrounding the active-site serine was Asn-Gly-Gly-Ser-Asn-Gly-Gly. This sequence is different from the active-site sequences of other known serine proteases. This difference and the lack of overall homology with the known families of serine proteases suggest that prolyl endopeptidase represents a new type of serine protease.
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页码:2195 / 2203
页数:9
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