Elkhatib Walid F, Hair Pamela S, Nyalwidhe Julius O, Cunnion Kenji M
Department of Pharmacy Practice, School of Pharmacy, Hampton University, Kittrell Hall, Hampton, VA 23668, USA.
Department of Pediatrics, Eastern Virginia Medical School, E.V. Williams Hall, 855 W. Brambleton Avenue, Norfolk, VA 23510, USA.
J Med Microbiol. 2015 Apr;64(Pt 4):335-343. doi: 10.1099/jmm.0.000010.
Staphylococcus aureus is a crucial human pathogen expressing various immune-evasion proteins that interact with the host-cell molecules. Clumping factor A (ClfA) is a microbial surface protein that promotes S. aureus binding to fibrinogen, and is associated with septic arthritis and infective endocarditis. In order to identify the major human serum proteins that bind the ClfA, we utilized recombinant ClfA region A in a plate-based assay. SDS-PAGE analysis of the bound proteins yielded five prominent bands, which were analysed by MS yielding apolipoprotein E (ApoE) as the predominant protein. ClfA-sufficient S. aureus bound purified ApoE by more than one log greater than an isogenic ClfA-deficient mutant. An immunodot-blot assay yielded a linearity model for ClfA binding to human ApoE with a stoichiometric-binding ratio of 1.702 at maximal Pearson's correlation coefficient (0.927). These data suggest that ApoE could be a major and novel binding target for the S. aureus virulence factor ClfA. Thus, ClfA recruitment of serum ApoE to the S. aureus surface may sequester ApoE and blunt its host defence function against S. aureus-invasive infections to humans. In this context, compounds that can block or suppress ClfA binding to ApoE might be utilized as prophylactic or therapeutic agents.
金黄色葡萄球菌是一种重要的人类病原体,可表达多种与宿主细胞分子相互作用的免疫逃避蛋白。凝聚因子A(ClfA)是一种微生物表面蛋白,可促进金黄色葡萄球菌与纤维蛋白原结合,并与脓毒性关节炎和感染性心内膜炎相关。为了鉴定与ClfA结合的主要人血清蛋白,我们在基于平板的检测中使用了重组ClfA区域A。对结合蛋白进行SDS-PAGE分析产生了五条明显的条带,通过质谱分析得出载脂蛋白E(ApoE)是主要蛋白。与同基因ClfA缺陷型突变体相比,表达ClfA的金黄色葡萄球菌结合纯化的ApoE的能力高出一个对数以上。免疫斑点印迹分析得出ClfA与人ApoE结合的线性模型,在最大皮尔逊相关系数(0.927)时化学计量结合比为1.702。这些数据表明,ApoE可能是金黄色葡萄球菌毒力因子ClfA的主要新型结合靶点。因此,ClfA将血清ApoE募集到金黄色葡萄球菌表面可能会隔离ApoE并削弱其对人类金黄色葡萄球菌侵袭性感染的宿主防御功能。在这种情况下,能够阻断或抑制ClfA与ApoE结合的化合物可作为预防或治疗药物。