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首页> 外文期刊>Infection and immunity >Identification and Molecular Analysis of Rough-Colony-Specific Outer Membrane Proteins of Actinobacillus actinomycetemcomitans
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Identification and Molecular Analysis of Rough-Colony-Specific Outer Membrane Proteins of Actinobacillus actinomycetemcomitans

机译：放线放线杆菌的粗集落特异外膜蛋白的鉴定和分子分析

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Actinobacillus actinomycetemcomitans, a gram-negative bacterium isolated from the human mouth, has been implicated in the pathogenesis of early-onset periodontitis. Primary isolates cultured from subgingival plaque exhibit an adherent, rough colony phenotype which spontaneously converts to a nonadherent, smooth phenotype upon in vitro subculture. The rough colony variant produces abundant fimbriae and autoaggregates, while the smooth colony variant is planktonic and produces scant fimbriae. To begin to understand the significance of colony variation in biofilm formation by A. actinomycetemcomitans, outer membrane protein profiles of four isogenic rough and smooth colony variants were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two proteins with relative molecular masses of 43 and 20 kDa were expressed by the rough colony variants exclusively. Expression of these proteins was not found to be dependent on growth phase, oxygen tension, or type of complex medium. N-terminal amino acid sequences of these proteins obtained by Edman degradation were compared with sequences from the University of Oklahoma A. actinomycetemcomitans genome database. Two contiguous open reading frames (ORFs) encoding proteins having sequence homology with these proteins were identified. The 43-kDa protein (RcpA [rough colony protein A]) was similar to precursor protein D of the general secretion pathway of gram-negative bacilli, while the 20-kDa protein (RcpB [rough colony protein B]) appeared to be unique. The genes encoding these proteins have been cloned from A. actinomycetemcomitans 283 and sequenced. A BLASTX (gapped BLAST) search of the surrounding ORFs revealed homology with other fimbria-related proteins. These data suggest that the genes encoding the 43-kDa (rcpA) and 20-kDa (rcpB) proteins may be functionally related to each other and to genes that may encode fimbria-associated proteins.
机译：从人口中分离出的革兰氏阴性细菌放线菌放线杆菌与早发性牙周炎的发病机制有关。从牙龈下斑块培养的主要分离株表现出粘附的粗糙菌落表型，在体外传代培养后可自发转化为非粘附的平滑表型。粗糙的菌落变体产生大量的菌毛并自动聚集，而光滑的菌落变体为浮游性且产生很少的菌毛。要开始了解菌落变异在 A生物膜形成中的重要性。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳比较了四个同基因的粗糙和光滑菌落变体的放线菌。相对分子质量分别为43和20 kDa的两种蛋白质仅通过粗糙集落变体表达。未发现这些蛋白质的表达取决于生长期，氧张力或复杂培养基的类型。将通过埃德曼降解获得的这些蛋白质的N-末端氨基酸序列与俄克拉何马大学的序列进行比较。放线菌基因组数据库。鉴定了两个连续的开放阅读框（ORF），它们编码与这些蛋白质具有序列同源性的蛋白质。 43-kDa蛋白（RcpA [粗糙集落蛋白A]）与革兰氏阴性杆菌一般分泌途径的前体蛋白D相似，而20-kDa蛋白（RcpB [粗糙集落蛋白B]）似乎是唯一的。 。已经从 A克隆了编码这些蛋白质的基因。放线放线菌 283并测序。 BLASTX（空白BLAST）搜索周围的ORF揭示了与其他菌毛相关蛋白的同源性。这些数据表明，编码43-kDa（ rcpA ）和20-kDa（ rcpB ）蛋白的基因可能在功能上相互关联，并且与可能编码菌毛的基因相关。相关蛋白。

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