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恶性疟原虫热稳定、菌株特异性S抗原的代谢标记与特性分析

Metabolic labelling and characterisation of S-antigens, the heat-stable, strain-specific antigens of Plasmodium falciparum.

作者信息

Winchell E J, Ling I T, Wilson R J

出版信息

Mol Biochem Parasitol. 1984 Mar;10(3):287-96. doi: 10.1016/0166-6851(84)90027-6.

Abstract

The C-10 clone of Plasmodium falciparum was metabolically labelled with [3H]glycine. Analysis by sodium dodecylsulphate/polyacrylamide gel electrophoresis (SDS/PAGE) revealed that S-antigens were the only significantly labelled products released into culture supernatants by the end of the asexual cycle. This result indicates conclusively that heat-stable, strain-specific antigens (the 'S-antigens') emanate from the parasite and not the host. The S-antigen of clone C-10 was resolved on SDS/PAGE as two labelled products with molecular weights of 156000 +/- 5000 and 130000 +/- 2000. The two components were differentially susceptible to proteolysis with trypsin. In other experiments, boiled plasma from Aotus monkeys infected with a different isolate of P. falciparum was partially purified by isoelectric focusing and the fraction containing S-antigen was iodinated by the Iodogen procedure. Analysis by SDS/PAGE of immunoprecipitated material revealed an iodinated component corresponding to the lower molecular weight band of the metabolically labelled S-antigen.

摘要

恶性疟原虫的C - 10克隆株用[³H]甘氨酸进行代谢标记。通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳(SDS/PAGE)分析表明,在无性生殖周期结束时,S抗原是释放到培养上清液中的唯一显著标记产物。这一结果确凿地表明,热稳定的、菌株特异性抗原(“S抗原”)源自疟原虫而非宿主。克隆C - 10的S抗原在SDS/PAGE上分离为两种标记产物,分子量分别为156000 ± 5000和130000 ± 2000。这两种成分对胰蛋白酶的蛋白水解作用敏感性不同。在其他实验中,用等电聚焦法对感染了不同恶性疟原虫分离株的夜猴的煮沸血浆进行部分纯化,并用碘代甘氨酸法对含有S抗原的部分进行碘化。对免疫沉淀物质进行SDS/PAGE分析,显示出一种碘化成分,对应于代谢标记的S抗原的较低分子量条带。

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