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首页> 外文期刊>Infection and immunity >Identification of a Staphylococcus aureus extracellular matrix-binding protein with broad specificity.
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Identification of a Staphylococcus aureus extracellular matrix-binding protein with broad specificity.

机译：具有广泛特异性的金黄色葡萄球菌细胞外基质结合蛋白的鉴定。

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A staphylococal surface protein capable of binding several extracellular matrix glycoproteins was purified as a result of our attempts to identify a receptor(s) for bone sialoprotein (BSP) on Staphylococcus aureus cells. Proteins from different staphylococcal strains were solubilized in sodium lauryl sulfate, separated by polyacrylamide gel electrophoresis, blotted onto Immobilon P membranes, and probed with 125I-BSP. Several bacterial proteins bound the radiolabeled ligand, and various strains expressed different repertoirs of BSP-binding proteins. Major BSP-binding proteins with apparent M(r)s of 72,000 or 60,000 were present on most strains, and these proteins were further studied. The 72- and 60-kDa proteins were preferentially expressed when bacteria were cultured in Luria broth compared with when they were cultured on tryptic soy broth, and the abundance of the proteins could be correlated to an increased 125I-BSP binding. Both the 72-kDa and the 60-kDa proteins were solubilized by extraction of cells with 1 M LiCl and were purified by cation-exchange chromatography. Amino acid composition analysis of the purified 72-kDa protein indicated a high content of lysine (11.9%) and hydrophobic amino acids (28.0% combined). In Western ligand blotting (immunoblotting) experiments, the 72-kDa protein bound not only BSP but also radiolabeled fibronectin, fibrinogen, vitronectin, thrombospondin, and, to some extent, collagen. Addition of the purified 60-kDa protein to S. aureus cells did not inhibit binding of the different ligands but in some cases resulted in an augmentation of the binding of 125I-ligand. Purified 60-kDa protein could hemagglutinate sheep erythrocytes at a concentration of 61 micrograms/ml. The agglutination reaction was inhibited by high concentrations of fucose, mannose, or melibiose. These data suggest that the purified proteins may serve as bacterial receptors with broad specificity for matrix glycoproteins and that the proteins may act as carbohydrate-binding proteins.
机译：由于我们试图鉴定金黄色葡萄球菌细胞上骨唾液蛋白（BSP）受体的结果，纯化了能够结合几种细胞外基质糖蛋白的葡萄球菌表面蛋白。将来自不同葡萄球菌菌株的蛋白质溶解在十二烷基硫酸钠中，通过聚丙烯酰胺凝胶电泳分离，印迹到Immobilon P膜上，并用125 I-BSP进行探测。几种细菌蛋白结合了放射性标记的配体，各种菌株表达了不同的BSP结合蛋白库。大多数菌株上均存在表观M（r）为72,000或60,000的主要BSP结合蛋白，并对这些蛋白进行了进一步研究。与在胰蛋白酶大豆肉汤中培养时相比，在Luria肉汤中培养细菌时优先表达72 kDa和60 kDa的蛋白，这些蛋白的丰度可能与125I-BSP结合的增加有关。通过用1 M LiCl提取细胞来溶解72 k​​Da和60 kDa的蛋白质，并通过阳离子交换色谱法进行纯化。纯化的72-kDa蛋白的氨基酸组成分析表明，赖氨酸（11.9％）和疏水性氨基酸（合计28.0％）含量高。在Western配体印迹（免疫印迹）实验中，72 kDa蛋白不仅结合BSP，而且还结合放射性标记的纤连蛋白，纤维蛋白原，玻连蛋白，血小板反应蛋白和某种程度上的胶原蛋白。向金黄色葡萄球菌细胞中添加纯化的60 kDa蛋白不会抑制不同配体的结合，但在某些情况下会导致125 I-配体结合的增加。纯化的60 kDa蛋白可以血凝集绵羊红细胞，浓度为61微克/毫升。高浓度的岩藻糖，甘露糖或蜜三糖可抑制凝集反应。这些数据表明，纯化的蛋白质可以用作对基质糖蛋白具有广泛特异性的细菌受体，并且这些蛋白质可以充当碳水化合物结合蛋白。

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