Access of antibody or trypsin to an integral outer membrane protein (P66) of Borrelia burgdorferi is hindered by Osp lipoproteins
Access of antibody or trypsin to an integral outer membrane protein (P66) of Borrelia burgdorferi is hindered by Osp lipoproteins
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DOI：
10.1128/iai.67.6.2874-2883.1999
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发表时间：
1999-06-01
影响因子：
3.1
通讯作者：
Barbour, AG
中科院分区：
文献类型：
--
作者：
Bunikis, J;Barbour, AG
莱姆病病原体伯氏疏螺旋体的外膜含有由其脂质部分锚定的脂蛋白和具有跨膜区域的整合蛋白。我们使用原位蛋白水解，免疫荧光，体外生长抑制，并与甲醛交联的技术来表征P66，一个完整的膜蛋白，和选择的Osp脂蛋白的B之间的拓扑关系。burgdorferi。蛋白酶处理的完整螺旋体裂解P66和Osp蛋白，但不是周质鞭毛蛋白或BmpA蛋白的细胞质膜，P66的细胞缺乏OspA，OspB，和OspC是更容易受到胰蛋白酶裂解比P66的细胞与这些Osp蛋白。抗P66表面环的单克隆抗体结合、凝集和抑制缺乏OspA、OspB、OspC和OspB的活螺旋体的生长，但不结合、凝集和抑制表达OspA、OspC和/或OspD的细胞的生长。当细胞被固定时，抗体结合表达OspD和OspC的细胞，但仍不结合具有OspA的细胞。通过甲醛交联两种蛋白质证实了这一点。这些结果表明，Osp蛋白，特别是OspA，限制抗体或胰蛋白酶接近P66的表面环区域。P66和OspA（或其他Osp蛋白）之间的接近和可能的接触可能会阻碍抗体对否则将是适当的疫苗靶标的抗体的有效性。
The outer membrane of Borrelia burgdorferi, the Lyme disease agent, contains lipoproteins anchored by their lipid moieties and integral proteins with membrane-spanning regions. We used the techniques of in situ proteolysis, immunofluorescence, in vitro growth inhibition, and cross-linking with formaldehyde to characterize topological relationships between P66, an integral membrane protein, and selected Osp lipoproteins of B. burgdorferi. Protease treatment of intact spirochetes cleaved P66 and Osp proteins but not the periplasmic flagellin or the BmpA protein of the cytoplasmic membrane, P66 of cells lacking OspA, OspB, and OspC was more susceptible to trypsin cleavage than was P66 of cells with these Osp proteins. A monoclonal antibody against the surface loop of P66 bound, agglutinated, and inhibited the growth of viable spirochetes lacking OspA, OspB, OspC, and OspB but not of the cells that expressed OspA, OspC, and/or OspD, When cells were fixed, the antibody bound to cells that express OspD and OspC but still not to cells with OspA, The close association of OspA and P66,vas confirmed by the crosslinking of the two proteins by formaldehyde. These results show that Osp proteins, particularly OspA, limit the access of antibody or trypsin to the surface loop region of P66. The proximity and possible contact between P66 and OspA (or other Osp proteins) may hinder the effectiveness of antibodies to what otherwise would be an appropriate vaccine target.