Patti J M, Jonsson H, Guss B, Switalski L M, Wiberg K, Lindberg M, Höök M
Department of Biochemistry, University of Alabama, Birmingham 35294-0005.
J Biol Chem. 1992 Mar 5;267(7):4766-72.
Some strains of Staphylococcus aureus bind collagen with a high degree of specificity and affinity. This interaction can represent a mechanism of substrate adhesion and may be an important step in the pathogenesis of osteomyelitis and infectious arthritis. We now report on the cloning, sequencing, and expression of a gene name cna, encoding a S. aureus collagen adhesin. The cna gene was isolated from a lambda GT11 S. aureus genomic library and encodes an 1185 amino acid polypeptide. The deduced amino acid sequence reveals several structural characteristics similar to previously described Gram-positive bacterial cell surface proteins. Antibodies raised against the native collagen adhesin from S. aureus recognize the recombinant collagen adhesin. Collagen binding activity can be detected in a lysate obtained from Escherichia coli cells, which harbor the cloned cna gene on an expression plasmid. Collagen-binding proteins can be detected in the lysate when analyzed by a Western blot type assay in which the membrane-transferred proteins are probed with radioactively labeled collagen. Finally, the bacterial lysate containing the recombinant adhesin can effectively inhibit the binding of soluble collagen to cells of S. aureus.
某些金黄色葡萄球菌菌株能高度特异性且亲和地结合胶原蛋白。这种相互作用可能代表一种底物黏附机制,并且可能是骨髓炎和感染性关节炎发病机制中的重要一步。我们现在报告一个名为cna的基因的克隆、测序及表达情况,该基因编码一种金黄色葡萄球菌胶原蛋白黏附素。cna基因是从λGT11金黄色葡萄球菌基因组文库中分离出来的,编码一个1185个氨基酸的多肽。推导的氨基酸序列显示出与先前描述的革兰氏阳性细菌细胞表面蛋白相似的几个结构特征。针对来自金黄色葡萄球菌的天然胶原蛋白黏附素产生的抗体能识别重组胶原蛋白黏附素。在从大肠杆菌细胞获得的裂解物中可检测到胶原蛋白结合活性,这些大肠杆菌细胞在表达质粒上携带克隆的cna基因。当通过蛋白质印迹类型分析(其中用放射性标记的胶原蛋白探测膜转移的蛋白质)进行分析时,在裂解物中可检测到胶原蛋白结合蛋白。最后,含有重组黏附素的细菌裂解物能有效抑制可溶性胶原蛋白与金黄色葡萄球菌细胞的结合。