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首页> 外文期刊>Infection and immunity >Sulfhydryl-dependent attachment of Treponema denticola to laminin and other proteins.
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Sulfhydryl-dependent attachment of Treponema denticola to laminin and other proteins.

机译：密螺旋体对密螺旋体和其他蛋白质的巯基依赖性附着。

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Attachment of Treponema denticola ATCC 35405 to laminin, a major basement membrane protein, and to other proteins was studied. Microdilution plates were coated with the proteins, and the attachment of T. denticola was measured by the enzyme-linked immunosorbent assay technique. Compared with bovine serum albumin (BSA), T. denticola had a high affinity to laminin, fibronectin, fibrinogen, and gelatin, as well as to type I and type IV collagens. Attachment to RGD peptide (Gly-Arg-Gly-Asp-Ser, the integrin recognition sequence) was only about 30% of that to laminin and was comparable to attachment to BSA. Tests with laminin fragments obtained through elastase digestion showed that the spirochetes attached well to an A-chain 140-kDa fragment involved in eukaryote cell attachment but did not attach to a 50-kDa fragment that includes the heparin binding site. Pretreatment of T. denticola with soluble laminin, fibronectin, gelatin, BSA, or fibrinogen had no effect on the attachment of the bacteria to laminin or fibronectin. A wide variety of compounds were tested for their possible inhibitory actions on the attachment. While most treatments of T. denticola ATCC 35405 had little or no effect on the attachment to proteins, sulfhydryl reagents p-chloromercuribenzoic acid (pCMBA) and oxidized glutathione inhibited the attachment by 70 to 99%, depending on the protein. When T. denticola was first allowed to attach to proteins, addition of pCMBA or oxidized glutathione could no longer reverse the attachment. Heat treatment of the spirochetes also markedly reduced the attachment to laminin, gelatin, and fibrinogen but not to BSA. Mixed glycosidase treatment of the spirochetes inhibited the attachment by 20 to 80%. None of the above treatments of the substrate proteins had any marked effect on the spirochete attachment. The results indicate that T. denticola has the capacity to bind to many different kinds of proteins by utilizing specific attachment mechanisms. The binding appears to involve protein SH groups and/or carbohydrate residues on the surface of T. denticola.
机译：研究了密闭螺旋体ATCC 35405与层粘连蛋白（一种主要的基底膜蛋白）和其他蛋白的连接。用蛋白质包被微量稀释板，并通过酶联免疫吸附测定技术测量T. denticola的附着。与牛血清白蛋白（BSA）相比，树状锥虫对层粘连蛋白，纤连蛋白，纤维蛋白原和明胶以及I型和IV型胶原蛋白具有很高的亲和力。与RGD肽的连接（Gly-Arg-Gly-Asp-Ser，整联蛋白识别序列）仅为层粘连蛋白的约30％，与BSA的连接相当。对通过弹性蛋白酶消化获得的层粘连蛋白片段进行的测试表明，螺旋体与附着在真核生物细胞附着中的A链140-kDa片段附着良好，但不附着于包含肝素结合位点的50-kDa片段。用可溶性层粘连蛋白，纤连蛋白，明胶，BSA或纤维蛋白原预处理锥虫对细菌与层粘连蛋白或纤连蛋白的附着没有影响。测试了多种化合物对附着的可能抑制作用。尽管大多数的T. denticola ATCC 35405处理方法对蛋白质的附着几乎没有影响，但是，巯基试剂对氯mercuribenzoic酸（pCMBA）和氧化型谷胱甘肽抑制了70％至99％的附着，具体取决于蛋白质。当首次允许T. denticola附着于蛋白质时，添加pCMBA或氧化型谷胱甘肽不再能破坏附着。螺旋体的热处理还显着减少了对层粘连蛋白，明胶和纤维蛋白原的附着，但对BSA的附着没有。螺旋体的混合糖苷酶处理将附着抑制了20％至80％。底物蛋白的上述处理均未对螺环蛇的附着产生任何显着影响。结果表明，T。denticola具有利用特异性附着机制与许多不同种类的蛋白质结合的能力。结合似乎涉及树形锥虫表面上的蛋白质SH基团和/或碳水化合物残基。

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