MOLECULAR CHARACTERIZATION OF PCP, THE STRUCTURAL GENE ENCODING THE PYRROLIDONE CARBOXYLYL PEPTIDASE FROM STREPTOCOCCUS-PYOGENES

被引:22
作者
CLEUZIAT, P [1 ]
AWADE, A [1 ]
ROBERTBAUDOUY, J [1 ]
机构
[1] INST NATL SCI APPL,GENET MOLEC MICROORGANISMES LAB,F-69621 VILLEURBANNE,FRANCE
关键词
D O I
10.1111/j.1365-2958.1992.tb01378.x
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
This paper describes the cloning of a gene (pcp) coding for pyrrolidone carboxylyl peptidase (PYRase), an enzyme which selectively removes N-terminal pyroglutamic acid residues from polypeptides. This gene was isolated from Streptococcus pyogenes by construction of a gene library with a bacteriophage lambda-derived cosmid-Escherichia coli host system. Nucleotide sequence determination of a 1.3 kb restriction fragment revealed a 645 bp open reading frame encoding a 215-amino-acid product of M(r)23135 consistent with the 26 kDa polypeptide obtained from in vivo overexpression in E. coli. Southern hybridization confirmed that pcp is a single-copy gene on the S. pyogenes chromosome. 5' and 3' endpoint mapping of the 0.7 kb specific transcript observed by Northern analysis permitted the identification of transcriptional initiation and termination signals. Structural features of the pcp gene product from S. pyogenes are discussed and compared with that from Bacillus subtilis. The lack of sequence identity with any other known protein or nucleotide sequence suggests that this enzyme belongs to a new class of peptidase.
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页码:2051 / 2063
页数:13
相关论文
共 78 条
[1]  
ALBERT Z, 1972, ACTA HISTOCHEM, V44, P98
[2]  
[Anonymous], [No title captured]
[3]   PYRROLIDONECARBOXYLYL PEPTIDASE - STABILIZATION AND PURIFICATION [J].
ARMENTROUT, RW ;
DOOLITTLE, RF .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1969, 132 (01) :80-+
[4]  
Ausubel FM, 1987, CURRENT PROTOCOLS MO
[5]   LINKAGE MAP OF ESCHERICHIA-COLI K-12, EDITION-8 [J].
BACHMANN, BJ .
MICROBIOLOGICAL REVIEWS, 1990, 54 (02) :130-197
[6]   BACILLUS-SUBTILIS REQUIRES A STRINGENT SHINE-DALGARNO REGION FOR GENE-EXPRESSION [J].
BAND, L ;
HENNER, DJ .
DNA-A JOURNAL OF MOLECULAR & CELLULAR BIOLOGY, 1984, 3 (01) :17-21
[7]  
BARTLER T, 1987, ARCH INTERN MED, V148, P1421
[8]  
BAUER K, 1981, EUR J BIOCHEM, V173, P173
[9]  
BLOMBACK B, 1967, METHOD ENZYMOL, V11, P398
[10]   STRUCTURE AND NUCLEOTIDE-SEQUENCE OF THE BACILLUS-SUBTILIS PHENYLALANYL-TRANSFER RNA-SYNTHETASE GENES [J].
BRAKHAGE, AA ;
WOZNY, M ;
PUTZER, H .
BIOCHIMIE, 1990, 72 (10) :725-734