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在分离的猪心脏瓣膜细胞上鉴定金黄色葡萄球菌结合蛋白。

Identification of Staphylococcus aureus binding proteins on isolated porcine cardiac valve cells.

作者信息

Campbell K M, Johnson C M

机构信息

Department of Pediatrics, Mayo Clinic, Rochester, MN 55905.

出版信息

J Lab Clin Med. 1990 Feb;115(2):217-23.

PMID:2299265
Abstract

Infective endocarditis caused by Staphylococcus aureus may be initiated by bacterial binding to cardiac valve cells. We investigated binding of whole S. aureus organisms to preparations of isolated porcine cardiac valve proteins. Cultured endothelial and subendothelial cells were surface labeled with iodine 125. After preabsorption with Escherichia coli, an organism that only rarely causes infective endocarditis, binding of surface proteins to S. aureus was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subsequent autoradiography. The results showed that cardiac valve endothelial cells expressed a major S. aureus-binding protein with an approximate apparent molecular weight of 120,000. In contrast, cardiac valve subendothelial cells expressed on their surface a single species of binding protein with an approximate apparent molecular weight of 220,000; immunoblot analysis suggested that this protein was fibronectin. We also used radiolabeled S. aureus to probe cellular proteins transferred to nitrocellulose membranes. This technique identified a 125,000 molecular weight protein that bound S. aureus in endothelial cell extracts. We conclude that specific S. aureus binding to cardiac valve cells is mediated by different receptors for endothelial and subendothelial cells.

摘要

金黄色葡萄球菌引起的感染性心内膜炎可能始于细菌与心脏瓣膜细胞的结合。我们研究了完整的金黄色葡萄球菌与分离的猪心脏瓣膜蛋白制剂的结合情况。培养的内皮细胞和内皮下细胞用碘125进行表面标记。在用很少引起感染性心内膜炎的大肠杆菌进行预吸收后,通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳和随后的放射自显影来评估表面蛋白与金黄色葡萄球菌的结合。结果显示,心脏瓣膜内皮细胞表达一种主要的金黄色葡萄球菌结合蛋白,其表观分子量约为120,000。相比之下,心脏瓣膜内皮下细胞在其表面表达一种单一的结合蛋白,表观分子量约为220,000;免疫印迹分析表明该蛋白是纤连蛋白。我们还使用放射性标记的金黄色葡萄球菌来探测转移到硝酸纤维素膜上的细胞蛋白。该技术鉴定出一种分子量为125,000的蛋白,它在内皮细胞提取物中与金黄色葡萄球菌结合。我们得出结论,金黄色葡萄球菌与心脏瓣膜细胞的特异性结合是由内皮细胞和内皮下细胞的不同受体介导的。

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