PURIFICATION AND CHARACTERIZATION OF PROLYL ENDOPEPTIDASE FROM PIG BRAIN

被引：18

作者：

SCHONLEIN, C ^{[1
]}

HEINS, J ^{[1
]}

BARTH, A ^{[1
]}

机构：

[1] UNIV HALLE SAALE, BIOTECHNIKUM, WEINBERGWEG 16A, O-4050 HALLE, GERMANY

来源：

BIOLOGICAL CHEMISTRY HOPPE-SEYLER | 1990年 / 371卷 / 12期

关键词：

PROLYL ENDOPEPTIDASE; PROTEIN PURIFICATION; PROTEIN MODIFICATION;

D O I：

10.1515/bchm3.1990.371.2.1159

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：

The prolyl endopeptidase from pig brain was purified to homogeneity according to SDS-gel electrophoresis and visualization with the silver staining procedure. The molecular weight of prolyl endopeptidase was estimated as 70 kDa, and the isoelectric point as 4.9. The molecular properties of prolyl endopeptidase from pig brain are therefore similar to those of prolyl endopeptidases from other mammalian tissues. Diisopropylfluorophosphate, diethylpyrocarbonate and p-chloromercuribenzoic acid are strong irreversible inhibitors of prolyl endopeptidase from pig brain. We showed that diisopropylfluorophosphate und diethylpyrocarbonate act as competitive inhibitors with respect to substrate. Therefore it is assumed that at least one serine and one histidine residue are located at the active site of this enzyme. This result supports the assumption that the prolyl endopeptidase from pig brain is a typical serine protease. Substance P, thyreoliberin, beta-casomorphin-5 and morphiceptin are hydrolysed by prolyl endopeptidase in vitro.

引用

页码：1159 / 1164

页数：6

共 27 条

[1] CHARACTERIZATION OF PROLINE ENDOPEPTIDASE FROM RAT-BRAIN [J].

ANDREWS, PC ;

HINES, CM ;

DIXON, JE .

BIOCHEMISTRY, 1980, 19 (24) :5494-5500

[2] IMPROVED SILVER STAINING OF PLANT-PROTEINS, RNA AND DNA IN POLYACRYLAMIDE GELS [J].

BLUM, H ;

BEIER, H ;

GROSS, HJ .

ELECTROPHORESIS, 1987, 8 (02) :93-99

[3] CLEAVAGE OF SUBSTANCE-P TO AN N-TERMINAL TETRAPEPTIDE AND A C-TERMINAL HEPTAPEPTIDE BY A POST-PROLINE CLEAVING ENZYME FROM BOVINE BRAIN [J].

BLUMBERG, S ;

TEICHBERG, VI ;

CHARLI, JL ;

HERSH, LB ;

MCKELVY, JF .

BRAIN RESEARCH, 1980, 192 (02) :477-486

[4]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[5] ESTIMATION OF MICHAELIS CONSTANT AND MAXIMUM VELOCITY FROM DIRECT LINEAR PLOT [J].

CORNISHBOWDEN, A ;

EISENTHAL, R .

BIOCHIMICA ET BIOPHYSICA ACTA, 1978, 523 (01) :268-272

[6] THE CONFORMATION AROUND THE PEPTIDE-BOND BETWEEN THE P1-POSITIONS AND P2-POSITIONS IS IMPORTANT FOR CATALYTIC ACTIVITY OF SOME PROLINE-SPECIFIC PROTEASES [J].

FISCHER, G ;

HEINS, J ;

BARTH, A .

BIOCHIMICA ET BIOPHYSICA ACTA, 1983, 742 (03) :452-462

[7] PROTEOLYTIC ENZYMES [J].

HARTLEY, BS .

ANNUAL REVIEW OF BIOCHEMISTRY, 1960, 29 :45-72

[8]

HARTRODT B, 1982, PHARMAZIE, V37, P72

[9]

JANSEN EF, 1949, J BIOL CHEM, V179, P189

[10]

KNISATSCHEK H, 1979, J BIOL CHEM, V254, P936

← 1 2 3 →