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首页> 外文期刊>Infection and immunity >Characterization, genetic analysis, and expression of a protease antigen (PrpRI) of Porphyromonas gingivalis W50.
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Characterization, genetic analysis, and expression of a protease antigen (PrpRI) of Porphyromonas gingivalis W50.

机译：牙龈卟啉单胞菌W50蛋白酶抗原（PrpRI）的表征，遗传分析和表达。

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Previous studies of the serum immunoglobulin G antibody response of periodontal patients have demonstrated significant reactivity to a cell surface or extracellular arginine-specific protease of Porphyromonas gingivalis which migrates as an approximately 50-kDa band on sodium dodecyl sulfate-polyacrylamide gels. In the present report, two forms of the enzyme (ArgI and ArgIA) with this electrophoretic behavior were isolated. ArgI is a heterodimer of alpha and beta subunits, and ArgIA is a monomer composed of the catalytically active alpha component alone. The gene encoding ArgI (prpR1 encoding protease polyprotein ArgI) was cloned from Sau3AI digests of P. gingivalis W50 DNA into pUC18. Sequence analysis demonstrated that the alpha and beta components are contiguous on the initial translation product and are flanked by large N- and C-terminal extensions. prpR1 is 97.5% identical to the rgp-1 gene from P. gingivalis H66. prpR1 expression in Escherichia coli demonstrated the presence of an internal transcription-translation initiation site which could permit independent expression of different regions of the polyprotein. Immunochemical analysis of P. gingivalis mid-logarithmic-phase cultures suggested that the processing of PrpRI may be closely coupled to its synthesis, with only the final stages taking place at the cell surface. Southern hybridization studies demonstrated that the prpR1 gene is widely distributed in other P. gingivalis strains and that a second homologous locus to the alpha component and at least two other homologous loci to the beta component are present on the P. gingivalis chromosome. These data indicate that the ArgI protease of P. gingivalis is a member of a family of sequence-related gene products which may share both functional and antigenic properties.
机译：牙周患者血清免疫球蛋白G抗体反应的先前研究表明，其对牙龈卟啉单胞菌的细胞表面或胞外精氨酸特异性蛋白酶具有显着反应性，该蛋白酶在十二烷基硫酸钠-聚丙烯酰胺凝胶上以大约50kDa的条带迁移。在本报告中，分离了具有这种电泳行为的两种形式的酶（ArgI和ArgIA）。 ArgI是α和β亚基的异二聚体，而ArgIA是仅由具有催化活性的α组分组成的单体。从齿龈假单胞菌W50 DNA的Sau3AI消化物中将编码ArgI的基因（编码蛋白酶多蛋白ArgI的prpR1）克隆到pUC18中。序列分析表明，α和β成分在初始翻译产物上是连续的，并在N和C末端有较大的延伸。 prpR1与牙龈卟啉单胞菌H66的rgp-1基因有97.5％的同一性。大肠杆菌中prpR1的表达证明存在内部转录翻译起始位点，该位点可允许独立表达多蛋白的不同区域。牙龈卟啉单胞菌对数中期培养的免疫化学分析表明，PrpRI的加工过程可能与其合成密切相关，只有最后阶段发生在细胞表面。 Southern杂交研究表明，prpR1基因广泛分布在其他齿龈假单胞菌菌株中，并且在齿龈假单胞菌染色体上存在与α成分的第二同源基因座和与β成分的至少两个其他同源基因座。这些数据表明牙龈卟啉单胞菌的ArgI蛋白酶是序列相关基因产物家族的成员，其可以共享功能和抗原特性。

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