SULFHYDRYL-DEPENDENT ATTACHMENT OF TREPONEMA-DENTICOLA TO LAMININ AND OTHER PROTEINS
SULFHYDRYL-DEPENDENT ATTACHMENT OF TREPONEMA-DENTICOLA TO LAMININ AND OTHER PROTEINS
复制标题
DOI：
10.1128/iai.59.11.4230-4237.1991
复制
发表时间：
1991-11-01
影响因子：
3.1
通讯作者：
UITTO, VJ
中科院分区：
文献类型：
--
作者：
HAAPASALO, M;SINGH, U;UITTO, VJ
研究了密螺旋体 ATCC 35405 与层粘连蛋白（一种主要基底膜蛋白）和其他蛋白的附着。将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齿垢毛细菌的附着情况。与牛血清白蛋白 (BSA) 相比，T. denticola 对层粘连蛋白、纤连蛋白、纤维蛋白原和明胶以及 I 型和 IV 型胶原蛋白具有较高的亲和力。 RGD 肽（Gly-Arg-Gly-Asp-Ser，整联蛋白识别序列）的附着量仅为层粘连蛋白的约 30%，与 BSA 的附着量相当。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，螺旋体很好地附着在参与真核细胞附着的 A 链 140 kDa 片段上，但没有附着在包含肝素结合位点的 50 kDa 片段上。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 T. denticola 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。虽然大多数 T. denticola ATCC 35405 处理对蛋白质的附着影响很小或没有影响，但巯基试剂对氯汞苯甲酸 (pCMBA) 和氧化型谷胱甘肽可抑制附着 70% 至 99%，具体取决于蛋白质。当 T. denticola 第一次被允许附着到蛋白质上时，添加 pCMBA 或氧化型谷胱甘肽不再能逆转这种附着。螺旋体的热处理也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 的附着没有影响。螺旋体的混合糖苷酶处理抑制了 20% 至 80% 的附着。上述底物蛋白的处理均未对螺旋体附着产生任何显着影响。结果表明，T. denticola 能够利用特定的附着机制与许多不同种类的蛋白质结合。这种结合似乎涉及 T. denticola 表面上的蛋白质 SH 基团和/或碳水化合物残基。
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.