ANTIGENIC RELATEDNESS AND N-TERMINAL SEQUENCE HOMOLOGY DEFINE 2 CLASSES OF PERIPLASMIC FLAGELLAR PROTEINS OF TREPONEMA-PALLIDUM SUBSP PALLIDUM AND TREPONEMA-PHAGEDENIS
ANTIGENIC RELATEDNESS AND N-TERMINAL SEQUENCE HOMOLOGY DEFINE 2 CLASSES OF PERIPLASMIC FLAGELLAR PROTEINS OF TREPONEMA-PALLIDUM SUBSP PALLIDUM AND TREPONEMA-PHAGEDENIS
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DOI：
10.1128/jb.170.9.4072-4082.1988
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发表时间：
1988-09-01
影响因子：
3.2
通讯作者：
LIMBERGER, RJ
中科院分区：
文献类型：
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作者：
NORRIS, SJ;CHARON, NW;LIMBERGER, RJ
许多螺旋体的周质鞭毛含有多种蛋白质。在这项研究中，双向电泳，蛋白质印迹（免疫印迹），免疫过氧化物酶染色，和N-末端氨基酸序列分析的特点，梅毒螺旋体亚种的个别周质鞭毛蛋白。pallidum（Nichols株）和T. 5. history纯化T. pallidum周质鞭毛含有6种蛋白（Mrs = 37，000、34，500、33，000、30，000、29，000和27，000），而T. phagedenis周质鞭毛含有一个主要的39，000-Mr蛋白和一组两个主要和两个次要的33，000-至34，000-Mr多肽种类，37，000-和30，000-Mr蛋白也存在于一些T.噬菌体制剂。单特异性抗血清和单克隆抗体的免疫印迹和N-末端序列分析表明，主要的周质鞭毛蛋白分为两个不同的类，指定为A类和B类。A类蛋白由T. pallidum和T. B类包括T. pallidum 34.5-、33-和30-kDa蛋白以及T.噬菌体每个类别内的蛋白质具有免疫交叉反应性，并具有相似的N-末端序列（67 - 95%同源性）;两个类别之间没有明显的交叉反应性或序列同源性。抗A类或抗B类抗体不与T. pallidum或37和30-kDa T.噬菌体蛋白，表明这些蛋白与A类和B类蛋白抗原性无关。每个生物体的主要周质鞭毛蛋白之间缺乏完整的N-末端序列同源性表明它们最有可能由单独的结构基因编码。此外，还对T. phagedenis和T.苍白球周质鞭毛蛋白是高度保守的，尽管这两个物种的遗传差异。
The periplasmic flagella of many spirochetes contain multiple proteins. In this study, two-dimensional electrophoresis, Western blotting (immunoblotting), immunoperoxidase staining, and N-terminal amino acid sequence anaylsis were used to characterize the individual periplasmic flagellar proteins of Trepontema pallidum subsp. pallidum (Nichols strain) and T. phagedenis Karzan 5. Purified T. pallidum periplasmic flagella contained six proteins (Mrs = 37,000, 34,500, 33,000, 30,000, 29,000, and 27,000), whereas T. phagedenis periplasmic flagella contained a major 39,000-Mr protein and a group of two major and two minor 33,000- to 34,000-Mr polypeptide species; 37,000- and 30,000-Mr proteins were also present in some T. phagedenis preparations. Immunoblotting with monospecific antisera and monoclonal antibodies and N-terminal sequence analysis indicate that the major periplasmic flagellar proteins were divided into two distinct classes, designated class A and class B. Class A proteins consisted of the 37-kilodalton (kDa) protein of T. pallidum and the 39-kDa polypeptide of T. phagedenis; class B included the T. pallidum 34.5-, 33-, and 30-kDa proteins and the four 33- and 34-kDa polypeptide species of T. phagedenis. The proteins within each class were immunologically cross-reactive and possessed similar N-terminal sequences (67 to 95% homology); no cross-reactivity or sequence homology was evident between the two classes. Anti-class A or anti-class B antibodies did not react with the 29- or 27-kDa polypeptides of T. pallidum or the 37- and 30-kDa T. phagedenis proteins, indicating that these proteins are antigenically unrelated to the class A and class B proteins. The lack of complete N-terminal sequence homology among the major periplasmic flagellar proteins of each organism indicates that they are most likely encoded by separate structural genes. Furthermore, the N-terminal sequences of T. phagedenis and T. pallidum periplasmic flagellar proteins are highly conserved, despite the genetic dissimilarity of these two species.