Identification of enolase as a laminin-binding protein on the surface of Staphylococcus aureus
Identification of enolase as a laminin-binding protein on the surface of Staphylococcus aureus
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DOI：
10.1016/j.micinf.2004.02.003
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发表时间：
2004-05-01
影响因子：
5.8
通讯作者：
Brentani, RR
中科院分区：
文献类型：
--
作者：
Carneiro, CRW;Postol, E;Brentani, RR
我们以前已经证明，金黄色葡萄球菌，一种高度侵袭性的细菌，提出了一个52 kDa的表面蛋白，介导其结合层粘连蛋白。为了更好地表征这种受体，我们从二维（2-D）PAGE中切下这种假定的层粘连蛋白受体，并将其用作抗原，用于产生用于筛选S. aureus表达文库。获得了一个0.3kb的克隆，其序列与S.金黄色葡萄球菌α-烯醇化酶。此外，从2-D凝胶洗脱的52-kDa蛋白质的氨基酸测序表明其与a-烯醇化酶的分子同源性，a-烯醇化酶是一种在物种之间呈现高度进化保守性的酶。与此同时，针对S.在Western印迹分析中，金黄色葡萄球菌52-kDa条带也识别酵母α-烯醇酶。这些单克隆抗体也能够促进捕获碘标记的细菌吸附到固相时，这种捕获被抑制过量的兔肌肉α-烯醇化酶的加入。最后，对S.通过流式细胞术进一步证实金黄色葡萄球菌α-烯醇酶。因此，α-烯醇化酶可能在链球菌的发病机制中起重要作用。金黄色葡萄球菌通过允许其粘附到含有层粘连蛋白的细胞外基质。(C)2004年，Elsevier SAS。All rights reserved.
We have previously demonstrated that Staphylococcus aureus, a highly invasive bacteria, presents a 52-kDa surface protein that mediates its binding to laminin. In order to better characterize this receptor, we excised this putative laminin receptor from two-dimensional (2-D) PAGE and used it as antigen for raising a mouse hyperimmune serum which was for screening an S. aureus expression library. A single clone of 0.3 kb was obtained, and its sequence revealed 100% homology with S. aureus a-enolase. Moreover, amino acid sequencing of the 52-kDa protein eluted from the 2-D gel indicated its molecular homology with a-enolase, an enzyme that presents a high evolutionary conservation among species. In parallel, monoclonal antibodies raised against the S. aureus 52-kDa band also recognized yeast a-enolase in western blot analysis. These monoclonal antibodies were also able to promote capture of iodine-labeled bacteria when adsorbed to a solid phase, and this capture was inhibited by the addition of excess rabbit muscle a-enolase. Finally, the cell surface localization of S. aureus a-enolase was further confirmed by flow cytometry. Hence, a-enolase might play a critical role in the pathogenesis of S. aureus by allowing its adherence to laminin-containing extracellular matrix. (C) 2004 Elsevier SAS. All rights reserved.