A general procedure for the isolation of heat-shock proteins from periodontopathogenic bacteria

被引：14

作者：

Hinode, D ^{[1
]}

Grenier, D ^{[1
]}

Mayrand, D ^{[1
]}

机构：

[1] UNIV LAVAL, FAC MED DENTAIRE, GRP RECH ECOL BUCCALE, LAVAL, PQ G1K 7P4, CANADA

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 1996年 / 25卷 / 03期

基金：

英国医学研究理事会;

关键词：

heat-shock protein; immunoreactivity; purification; periodontopathogenic bacteria;

D O I：

10.1016/0167-7012(96)00008-5

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The isolation of heat-shock proteins (HSP) from whole cells of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Bacteroides forsythus, which are recognized as periodontopathogens, was performed by affinity chromatography on adenosine 5'-triphosphate-agarose followed by preparative polyacrylamide gel electrophoresis under denaturing conditions. Analysis of the final material by silver nitrate staining of SDS-PAGE gels and Western immunoblotting using a commercial polyclonal antibody against HSP60 from Synechococcus sp. revealed homogenous protein preparations corresponding to GroEL-like proteins with apparent molecular mass of 68 kDa (P. gingivalis), 64 kDa (A. actinomycetemcomitans) and 67 kDa (B. forsythus). A second HSP was also isolated from P. gingivalis and B. forsythus, and recognized as a DnaK-like protein by immunoblotting with polyclonal antibody against DnaK (HSP70) from Escherichia coli. Our HSP isolation procedure is rapid and simple, and the purified proteins obtained may be used to determine their homology with other HSP previously characterized and to raise antibodies.

引用

页码：349 / 355

页数：7

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