BmpA is a surface-exposed outer-membrane protein of Borrelia burgdorferi.
BmpA is a surface-exposed outer-membrane protein of Borrelia burgdorferi.
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DOI：
10.1111/j.1574-6968.2010.02020.x
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发表时间：
2010-08-01
影响因子：
2.1
通讯作者：
Cabello FC
中科院分区：
文献类型：
--
作者：
Bryksin AV;Tomova A;Godfrey HP;Cabello FC
Bmpa是伯氏疏螺旋体的一种免疫优势蛋白，也是一种致炎因子。制备兔抗重组BmpA(RBmpA)抗体，检测其与rBmpB、rBmpC和rBmpD的交叉反应性，然后通过与rBmpB的吸附使其具有单特异性。这种单特异性试剂只与rBmpA发生斑点免疫结合，并在二维免疫印迹上检测到单一的39 kDa的pI为5.0的斑点。用它来评估BMPA细胞的位置。在Triton X-114细胞的洗涤剂可溶组分和不溶组分中均存在Bmpa，提示它是一种膜脂蛋白。蛋白水解酶K处理的完整细胞和Triton X-100通透性细胞的免疫印迹显示，完整细胞中的BMPA被消化，与表面暴露一致。双重标记免疫荧光显微镜证实了这种暴露，细胞完好和通透性良好。Bmpa在所有伯氏杆菌基因组中的保守和表面定位可能表明它在该生物的生物学和病理生物学中发挥着关键作用。
BmpA is an immunodominant protein of Borrelia burgdorferi as well as an arthritogenic factor. Rabbit anti-recombinant BmpA (rBmpA) antibodies were raised, characterized by assaying their cross reactivity with rBmpB, rBmpC and rBmpD, 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 BmpA cellular location. BmpA was present in both detergent-soluble and -insoluble fractions of Triton X-114 phase-partitioned borrelial cells, suggesting 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.
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