The Mycoplasma gallisepticum α-enolase is cell surface-exposed and mediates adherence by binding to chicken plasminogen

被引：46

作者：

Chen, Hongjun ^{[2
]}

Yu, Shengqing ^{[1
,2
]}

Shen, Xinyue ^{[2
]}

Chen, Danqing ^{[2
]}

Qiu, Xvsheng ^{[2
]}

Song, Cuiping ^{[2
]}

Ding, Chan ^{[1
,2
]}

机构：

[1] China Natl Engn Technol Res Ctr Poultry, Shanghai 200331, Peoples R China

[2] Chinese Acad Agr Sci, Shanghai Vet Res Inst, Shanghai 200241, Peoples R China

来源：

MICROBIAL PATHOGENESIS | 2011年 / 51卷 / 04期

基金：

中国国家自然科学基金;

关键词：

Mycoplasma gallisepticum; alpha-Enolase; Plasminogen; Adherence; GENOME-REDUCED BACTERIUM; STREPTOCOCCUS-PNEUMONIAE; IDENTIFICATION; CYTADHESIN; PROTEIN; PATHOGENESIS; FERMENTANS; INVASION; ANTIGEN; SUIS;

D O I：

10.1016/j.micpath.2011.03.012

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

The alpha-enolase protein is reported to be an adhesin in several pathogenic bacterial species, but its role in Mycoplasma gallisepticum is unknown. In this study, the M. gallisepticum alpha-enolase gene was adapted to heterologous expression in Escherichia coli by performing overlapping polymerase chain reaction with site-directed mutagenesis to introduce A960G and A1158G mutations in the nucleotide sequence. The full-length mutated gene was cloned into a pGEM-T Easy vector and subcloned into the expression vector pET32a(+) to construct the pET-rMGEno plasmid. The expression of rMGEno in E. coli strain DE3 was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis with Coomassie blue staining. Purified rMGEno exhibited alpha-enolase catalytic activity that it could reflect the conversion of NADH to NAD(+). Mouse antiserum to alpha-enolase was generated by immunization with rMGEno. Immunoblotting and immunofluorescence assay with the antiserum identified alpha-enolase on the surface of M. gallisepticum cells. Enzyme-linked immunosorbent assay characterized rMGEno as a chicken plasminogen binding protein. An adherence inhibition assay on immortalized chicken fibroblasts (DF-1) demonstrated more than 77% inhibition of adhesion in the presence of mouse antiserum, suggesting that alpha-enolase of M. gallisepticum participates in bacterial adhesion to DF-1 cells. (C) 2011 Elsevier Ltd. All rights reserved.

引用

页码：285 / 290

页数：6

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