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Monoclonal antibodies that recognize a polypeptide antigenic determinant shared by multiple Caenorhabditis elegans sperm-specific proteins

机译:识别多种秀丽隐杆线虫精子特异性蛋白共有的多肽抗原决定簇的单克隆抗体

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摘要

Four monoclonal antibodies that are directed against antigens present in sperm and absent from other worm tissues were characterized. Antibody TR20 is directed against the major sperm proteins, a family of small, abundant, cytoplasmic proteins that have been previously described (Klass, M. R., and D. Hirsh, 1981, Dev. Biol., 84:299-312; Burke, D. J., and S. Ward, 1983, J. Mol. Biol., 171:1-29). Three other antibodies, SP56, SP150, and TR11, are all directed against the same set of minor sperm polypeptides that range in size from 29 to 215 kD. More than eight different sperm polypeptides are antigenic by both immunotransfer and immunoprecipitation assays. The three antibodies are different immunoglobulin subclasses, yet they compete with each other for antigen binding so they are directed against the same antigenic determinant on the multiple sperm proteins. This antigenic determinant is sensitive to any of six different proteases, is insensitive to periodate oxidation or N-glycanase digestion, and is detectable on a polypeptide synthesized in vitro. Therefore, the antigenic determinant resides in the polypeptide chain. However, peptide fragments of the proteins are not antigenic, thus the determinant is likely to be dependent on polypeptide conformation. The antigenic determinant shared by these proteins could represent a common structural feature of importance to the localization or cellular specificity of these proteins.
机译:鉴定了四种针对精子中存在的抗原以及其他蠕虫组织中不存在的抗原的单克隆抗体。抗体TR20针对主要的精子蛋白,这是一种先前描述的小而丰富的细胞质蛋白家族(Klass,MR,and D. Hirsh,1981,Dev。Biol。,84:299-312; Burke,DJ ,和S. Ward,1983,分子生物学杂志,171:1-29)。其他三种抗体SP56,SP150和TR11均针对大小范围为29至215 kD的同一组次要精子多肽。通过免疫转移和免疫沉淀测定,超过八种不同的精子多肽具有抗原性。这三种抗体是不同的免疫球蛋白亚类,但它们彼此竞争抗原结合,因此它们针对多个精子蛋白上的相同抗原决定簇。该抗原决定簇对六种不同蛋白酶中的任一种敏感,对高碘酸盐氧化或N-聚糖酶消化不敏感,并且可在体外合成的多肽上检测到。因此,抗原决定簇位于多肽链中。但是,蛋白质的肽片段不是抗原性的,因此决定因素可能取决于多肽的构象。这些蛋白共有的抗原决定簇可能代表了一个共同的结构特征,对这些蛋白的定位或细胞特异性具有重要意义。

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