Assembly and antigenicity of the Neisseria gonorrhoeae pilus mapped with antibodies
Assembly and antigenicity of the Neisseria gonorrhoeae pilus mapped with antibodies
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DOI：
10.1128/iai.64.2.644-652.1996
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发表时间：
1996-02-01
影响因子：
3.1
通讯作者：
Tainer, JA
中科院分区：
文献类型：
--
作者：
Forest, KT;Bernstein, SL;Tainer, JA
使用一组定点抗体探针并通过肽绘制抗菌毛抗血清的特异性，研究了淋病奈瑟菌菌毛蛋白序列与其四级组装成菌毛纤维之间的关系。通过竞争性免疫测定和免疫电子显微镜，使用针对跨越菌毛蛋白序列的 11 种肽的多克隆抗体，鉴定了组装药丸中埋藏和暴露的肽。菌毛不与菌毛蛋白亚基显着竞争结合针对残基 13 至 31 (13-31) 和 18-36 的抗体，菌毛纤维与菌毛蛋白亚基充分竞争结合针对肽 37-56、58-78、110-120、115-127、122-139 和 140-159 产生的抗体，并且针对残留物的抗体竞争较弱 79-93和94-108，序列保守残基37-56和半保守残基94-108的抗体优先结合菌毛末端，如免疫电子显微镜所示。通过针对代表来自淋病奈瑟菌 MS11 菌毛序列的所有可能的六聚或八聚肽的重叠肽组的抗血清特异性的肽图来测试菌毛区域对免疫系统的暴露。通过对 这些区域是在兔子、小鼠和人类的抗菌血清以及患有淋病的人类志愿者的血清中测量的。这三个物种之间抗原反应的保守性和变化阐明了其他物种的免疫学研究与人类针对病原体的免疫反应的相关性。总的来说，我们的结果解释了整个菌毛蛋白 N 端三分之一的极端保守性，因为它在菌毛组装中起主导作用：疏水性残基 1-36 涉及隐藏的横向接触，极性残基 37-56 涉及菌毛纤维内的纵向接触。
The relationship between the sequence of Neisseria gonorrhoeae pilin and its quaternary assembly into pilus fibers was studied with a set of site directed antibody probes and by mapping the specificities of antipilus antisera with peptides. Buried and exposed peptides in assembled pill were identified by competitive immunoassays and immunoelectron microscopy with polyclonal antibodies raised against 11 peptides spanning the pilin sequence. Pili did not compete significantly with pilin subunits for binding to antibodies against residues 13 to 31 (13-31) and 18-36, Pilus fibers competed well with pilin protein subunits for binding to antibodies raised against peptides 37-56, 58-78, 110-120, 115-127, 122-139, and 140-159 and competed weakly for antibodies against residues 79-93 and 94-108, Antibodies to sequence-conserved residues 37-56 and to semiconserved residues 94-108 preferentially bound pilus ends as shown by immunoelectron microscopy. The exposure of pilus regions to the immune system was tested by peptide mapping of antiserum specificities against sets of overlapping peptides representing all possible hexameric or octameric peptides from the N. gonorrhoeae MS11 pilin sequence, The immunogenicity of exposed peptides incorporating semiconserved residues 49-56 and 121-126 was revealed by strong, consistent antigenic reactivity to these regions measured in antipilus sera from rabbits, mice, and humans and in sera from human volunteers with gonorrhea. The conservation and variation of antigenic responses among these three species clarify the relevance of immunological studies of other species to the human immune response against pathogens. Overall, our results explain the extreme conservation of the entire N-terminal one-third of the pilin protein by its dominant role in pilus assembly: hydrophobic residues 1-36 are implicated in buried lateral contacts, and polar residues 37-56 are implicated in longitudinal contacts within the pilus fiber.