Staggs T M, Greer M K, Baseman J B, Holt S C, Tryon V V
Department of Microbiology, University of Texas Health Science Center at San Antonio 78284-7758.
Mol Microbiol. 1994 May;12(4):613-9. doi: 10.1111/j.1365-2958.1994.tb01048.x.
Lactoferrin-binding or -associated proteins were identified in Treponema pallidum subspecies pallidum and Treponema denticola by affinity column chromatography using human lactoferrin and detergent-solubilized, radiolabelled spirochaetes. Two discrete polypeptides of T. pallidum with masses of 45 and 40 kDa and a broad band from 29-34 kDa exhibited association with human apo- and partially ferrated lactoferrin. T. denticola produced two proteins that associated with a lactoferrin affinity matrix (50 and 35 kDa). T. pallidum and T. denticola did not associate with soluble, human transferrin in parallel experiments. Soluble human lactoferrin competed with all lactoferrin-associated proteins from T. pallidum and T. denticola in competitive-binding assays. However, the T. denticola proteins dissociated from a lactoferrin-affinity matrix in the presence of differing concentrations of unlabelled, soluble lactoferrin competitor. Treatment with phospholipase D altered migration of the diffuse 29-34 kDa band of T. pallidum suggesting that the polypeptide was lipid-modified. Each of the lactoferrin-binding proteins from T. pallidum and T. denticola reacted with pooled rabbit syphilitic antisera. The lactoferrin-binding proteins of T. pallidum reacted with human sera from patients at all stages of syphilis. In addition, a monoclonal antibody generated against the 45 kDa polypeptide of T. pallidum crossreacted with the 29-34 kDa protein.
通过使用人乳铁蛋白以及去污剂增溶的放射性标记螺旋体进行亲和柱色谱法，在梅毒螺旋体苍白亚种和齿垢密螺旋体中鉴定出乳铁蛋白结合或相关蛋白。梅毒螺旋体有两种离散的多肽，质量分别为45 kDa和40 kDa，还有一条29 - 34 kDa的宽带，它们与人脱铁乳铁蛋白和部分铁化乳铁蛋白表现出结合。齿垢密螺旋体产生了两种与乳铁蛋白亲和基质结合的蛋白（50 kDa和35 kDa）。在平行实验中，梅毒螺旋体和齿垢密螺旋体与可溶性人转铁蛋白不结合。在竞争结合试验中，可溶性人乳铁蛋白与梅毒螺旋体和齿垢密螺旋体的所有乳铁蛋白相关蛋白竞争。然而，在不同浓度未标记的可溶性乳铁蛋白竞争者存在的情况下，齿垢密螺旋体的蛋白从乳铁蛋白亲和基质上解离。用磷脂酶D处理改变了梅毒螺旋体29 - 34 kDa弥散带的迁移，表明该多肽被脂质修饰。梅毒螺旋体和齿垢密螺旋体的每种乳铁蛋白结合蛋白都与兔梅毒混合抗血清发生反应。梅毒螺旋体的乳铁蛋白结合蛋白与梅毒各阶段患者的血清发生反应。此外，针对梅毒螺旋体45 kDa多肽产生的单克隆抗体与29 - 34 kDa蛋白发生交叉反应。
