Department of Microbiology and Immunology, New York Medical College, Valhalla, NY 10595, USA.
FEMS Microbiol Lett. 2010 Aug 1;309(1):77-83. doi: 10.1111/j.1574-6968.2010.02020.x. Epub 2010 May 20.
BmpA is an immunodominant protein of Borrelia burgdorferi as well as an arthritogenic factor. Rabbit antirecombinant BmpA (rBmpA) antibodies were raised, characterized by assaying their cross reactivity with rBmpB, rBmpC and rBmpD, and then rendered monospecific by absorption with rBmpB. This monospecific reagent reacted only with rBmpA in dot immunobinding and detected a single 39 kDa, pI 5.0, spot on two-dimensional immunoblots. It was used to assess the BmpA cellular location. BmpA was present in both detergent-soluble and -insoluble fractions of Triton X-114 phase-partitioned borrelial cells, suggesting that it was a membrane lipoprotein. Immunoblots of proteinase K-treated intact and Triton X-100 permeabilized cells showed digestion of BmpA in intact cells, consistent with surface exposure. This exposure was confirmed by dual-label immunofluorescence microscopy of intact and permeabilized borrelial cells. Conservation and surface localization of BmpA in all B. burgdorferi sensu lato genospecies could point to its playing a key role in this organism's biology and pathobiology.
BmpA 是伯氏疏螺旋体的一种免疫优势蛋白，也是一种致关节炎因子。制备了兔抗重组 BmpA(rBmpA)抗体，通过测定其与 rBmpB、rBmpC 和 rBmpD 的交叉反应性进行了表征，然后用 rBmpB 吸收使其成为单特异性抗体。该单特异性试剂在 dot 免疫结合反应中仅与 rBmpA 反应，并在二维免疫印迹上检测到单个 39 kDa、pI 5.0 的斑点。它用于评估 BmpA 的细胞位置。BmpA 存在于 Triton X-114 相分区伯氏疏螺旋体细胞的去污剂可溶性和不可溶性部分中，表明它是一种膜脂蛋白。蛋白酶 K 处理的完整细胞和 Triton X-100 通透细胞的免疫印迹显示完整细胞中 BmpA 的消化，与表面暴露一致。这一暴露通过完整和通透的伯氏疏螺旋体细胞的双标记免疫荧光显微镜得到证实。BmpA 在所有伯氏疏螺旋体属亚种中的保守性和表面定位可能表明它在该生物体的生物学和病理生物学中发挥着关键作用。
