A surface-exposed region of a novel outer membrane protein (P66) of Borrelia spp. is variable in size and sequence
A surface-exposed region of a novel outer membrane protein (P66) of Borrelia spp. is variable in size and sequence
复制标题
DOI：
10.1128/jb.180.7.1618-1623.1998
复制
发表时间：
1998-04-01
影响因子：
3.2
通讯作者：
Barbour, AG
中科院分区：
文献类型：
--
作者：
Bunikis, J;Luke, CJ;Barbour, AG
建立了莱姆病疏螺旋体66 kDa外膜蛋白(P66)的模型，预测在C末端附近有一个表面暴露环，该区域含有莱姆病患者血清普遍识别的抗原。在本研究中，我们利用伯氏疏螺旋体p66的单抗、伯氏疏螺旋体p66的表位定位和DNA测序，进一步研究了p66和其他疏螺旋体的同源蛋白，以进一步研究该蛋白。克隆了与伯氏疏螺旋体p66同源蛋白的全长基因，推导的蛋白全长为589个氨基酸，与伯氏疏螺旋体p66的同源性为58%。根据保守序列设计了与伯氏杆菌p66蛋白相同的表面暴露区域，利用基于保守序列和聚合酶链式反应设计的引物，我们鉴定并克隆了斑点疏螺旋体、斑疏螺、珊瑚疏螺旋体和鹅疏螺p66蛋白的相同区域，所有物种的p66蛋白序列都在一个表面暴露环的两侧显示了两个保守的疏水区，不同的疏螺旋体之间的环序列在序列和大小上都有很大的差异，在35到45个氨基酸之间变化，尽管疏螺旋体的实际功能是35到45个氨基酸。是未知的，结果表明它的表面暴露区域受到选择压力。
A model of the 66-kDa outer membrane protein (P66) of Lyme disease Borrelia spp, predicts a surface-exposed loop near the C terminus, This region contains an antigen commonly recognized by sera from Lyme disease patients, In the present study, this region of P66 and homologous proteins of other Borrelia spp, were further investigated by using monoclonal antibodies, epitope mapping of P66 of Borrelia burgdorferi, and DNA sequencing, A monoclonal antibody specific for B. burgdorferi bound to the portion of P66 that was accessible to proteolysis in situ, The linear epitope for the antibody was mapped within a variable segment of the surface-exposed region, To further study this protein, the complete gene of Borrelia hermsii for a protein homologous to P66 was cloned, The deduced protein was 589 amino acids in length and 58% identical to P66 of B. burgdorferi. The B. hermsii P66 protein was predicted to have a surface-exposed region in the same location as that of B. burgdorferi's P66 protein, With primers designed on the basis of conserved sequences and PCR, we identified and cloned the same regions of P66 proteins of Borrelia turicatae, Borrelia parkeri, Borrelia coriaceae, and Borrelia anserina, The deduced protein sequences from all species demonstrated two conserved hydrophobic regions flanking a surface-exposed loop, The loop sequences were highly variable between different Borrelia spp, in both sequence and size, varying between 35 and 45 amino acids, Although the actual function of P66 of Borrelia spp. is unknown, the results suggest that its surface-exposed region is subject to selective pressure.