Analysis of polypeptide composition and antigenic components of Rickettsia tsutsugamushi by polyacrylamide gel electrophoresis and immunoblotting
Analysis of polypeptide composition and antigenic components of Rickettsia tsutsugamushi by polyacrylamide gel electrophoresis and immunoblotting
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聚丙烯酰胺凝胶电泳和免疫印迹分析恙虫病立克次体的多肽组成和抗原成分
DOI：
10.1128/iai.48.3.671-675.1985
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发表时间：
1985
影响因子：
3.1
通讯作者：
M. Oyanagi
中科院分区：
文献类型：
--
作者：
A. Tamura;N. Ohashi;H. Urakami;K. Takahashi;M. Oyanagi
对恙虫病立克次体纯化液进行聚丙烯酰胺凝胶电泳，发现30多条多肽带，其中分子大小分别为70、60、54 ~ 56和46 ~ 47千道尔顿。与Gilliam、Karp、Kato菌株和新分离的Shimokoshi菌株的立克次体多肽组成比较，Kato菌株（54-56K）和Karp菌株（46-47K）的主多肽迁移速度分别略快于其他菌株的主多肽迁移速度。最大的主要多肽（54-56K）可以被胰蛋白酶处理完整的立克次体消化，并且在不同的制剂中含量不同，这表明该多肽存在于立克次体表面，并且在处理这些微生物时容易降解。对完整立克次体进行放射性碘化处理，并对裂解液进行聚丙烯酰胺凝胶电泳，检测立克次体表面54-56K和46-47K多肽；然而，70K和60K多肽未标记。用豚鼠制备的高免疫血清对各菌株进行免疫印迹实验表明，70K、54-56K和46-47K多肽具有抗原性。54-56K多肽具有菌株特异性，而70K和46-47K多肽与异源抗血清发生交叉反应。
Polyacrylamide gel electrophoresis of lysates of purified Rickettsia tsutsugamushi revealed as many as 30 polypeptide bands, including major bands corresponding to molecular sizes of 70, 60, 54 to 56, and 46 to 47 kilodaltons. Compared with the polypeptide composition of the rickettsiae of Gilliam, Karp, and Kato strains and a newly isolated Shimokoshi strain, the major polypeptide in the Kato strain (54-56K) and in the Karp strain (46-47K) migrated a little faster and slower, respectively, than the corresponding polypeptides in the other strains. The largest major polypeptide (54-56K) was digestible by the treatment of intact rickettsiae with trypsin and variable in content in separate preparations, suggesting that the polypeptide exists on the rickettsial surface and is easily degraded during the handling of these microorganisms. Several surface polypeptides of rickettsiae, including the 54-56K and 46-47K polypeptides, were detected by radioiodination of intact rickettsiae followed by polyacrylamide gel electrophoresis of the lysate; however, the 70K and 60K polypeptides were not labeled. Immunoblotting experiments with hyperimmune sera prepared in guinea pigs against each strain demonstrated that the 70K, 54-56K, and 46-47K polypeptides showed antigenic activities. The 54-56K polypeptide appeared to be strain specific, whereas the 70K and 46-47K polypeptides cross-reacted with the heterologous antisera.