PURIFICATION AND CHARACTERIZATION OF A FIBRINOGENOLYTIC SERINE PROTEINASE FROM ASPERGILLUS-FUMIGATUS CULTURE FILTRATE

被引：92

作者：

LARCHER, G ^{[1
]}

BOUCHARA, JP ^{[1
]}

ANNAIX, V ^{[1
]}

SYMOENS, F ^{[1
]}

CHABASSE, D ^{[1
]}

TRONCHIN, G ^{[1
]}

机构：

[1] INST HYG & EPIDEMIOL, B-1050 BRUSSELS, BELGIUM

来源：

FEBS LETTERS | 1992年 / 308卷 / 01期

关键词：

ENZYME PURIFICATION; SERINE PROTEINASE; FIBRINOGENOLYTIC; ASPERGILLUS-FUMIGATUS;

D O I：

10.1016/0014-5793(92)81052-N

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：

A fibrinogenolytic proteinase has been isolated from Aspergillus fumigatus culture filtrate by ammonium sulfate precipitation followed by successive chromatographies on Sephadex G-75 and immobilized phenylalanine. The purified proteinase exhibited a molecular weight of about 33 kDa. When analysed by SDS-polyacrylamide gels containing co-polymerized fibrinogen, the proteinase appeared as a broad band at the top of the gels, which could correspond to polymerization of the enzyme, as suggested by SDS-PAGE analysis of the unboiled eluate. The isoelectric point was 8.75 and the enzyme was not glycosylated. Proteinase activity was optimum at pH 9 and between 37 and 42-degrees-C, although a decrease in activity was observed above 37-degrees-C. PMSF and chymostatin markedly inhibited the proteinase activity, and good kinetic constants were obtained for the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA. These results provide direct evidence that this enzyme belongs to the chymotrypsin-like serine proteinase group.

引用

页码：65 / 69

页数：5

共 26 条

[1] SPECIFIC BINDING OF HUMAN FIBRINOGEN FRAGMENT-D TO ASPERGILLUS-FUMIGATUS CONIDIA [J].