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黏菌原质团肌动蛋白抗体的诱导及其性质

Inducation of antibody against actin from myxomycete plasmodium and its properties.

作者信息

Owaribe K, Hatano S

出版信息

Biochemistry. 1975 Jul;14(13):3024-9. doi: 10.1021/bi00684a035.

Abstract

Plasmodium actin was highly purified by gel filtration of crude G-actin on Sephadex G-100 followed by ultracentrifugation after polymerization in the presence of 1 M urea and 1 mM ATP. Purified actin showed a single band in the sodium dodecyl sulfate gel electrophoretic pattern. Antibody against this purified actin was induced in rabbits. The antibody obtained was immunologically monospecific for plasmodium actin, judging from the following results. (1) The addition of the antibody to a plasmodium F-action solution increased the turbidity of the mixed solution, showing the formation of the antibody-action complex. (2) In immunodiffusion and immunoelectrophoresis, the antibody formed single preciptin lines with the purified actin preparation and with the crude actin extract from the acetone-dried powder of plasmodium. (3) The antibody inhibited polymerization of plasmodium G-actin. (4) Plasmodium F-actin filaments were decorated with antibody in electron micrographs. The antibody reacted not only with plasmodium F- and G-actin, but also reacted with sea urchin egg actin, but it did not react with actin from rabbit striated muscle.

摘要

通过在1 M尿素和1 mM ATP存在下聚合后,将粗制G -肌动蛋白在Sephadex G - 100上进行凝胶过滤,随后进行超速离心,从而高度纯化疟原虫肌动蛋白。纯化后的肌动蛋白在十二烷基硫酸钠凝胶电泳图谱中显示为单一条带。在兔体内诱导产生针对这种纯化肌动蛋白的抗体。从以下结果判断,所获得的抗体对疟原虫肌动蛋白具有免疫单特异性。(1)将该抗体添加到疟原虫F -肌动蛋白溶液中会增加混合溶液的浊度,表明形成了抗体 - 肌动蛋白复合物。(2)在免疫扩散和免疫电泳中,该抗体与纯化的肌动蛋白制剂以及来自疟原虫丙酮干粉的粗肌动蛋白提取物形成单一沉淀线。(3)该抗体抑制疟原虫G -肌动蛋白的聚合。(4)在电子显微镜照片中,疟原虫F -肌动蛋白丝被抗体标记。该抗体不仅与疟原虫F -肌动蛋白和G -肌动蛋白反应,还与海胆卵肌动蛋白反应,但不与兔横纹肌肌动蛋白反应。

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