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旋盘尾丝虫发育阶段中的磷蛋白模式

Phosphoprotein patterns in Onchocerca volvulus developmental stages.

作者信息

Muluh J P, Titanji V P, Zetterqvist O

机构信息

Biotechnology Centre, Yaoundé, Sweden.

出版信息

Ups J Med Sci. 1994;99(1):39-49. doi: 10.3109/03009739409179349.

Abstract

Living females and microfilariae of Onchocerca volvulus incubated in culture medium containing [32P]orthophosphate were observed to phosphorylate their proteins rapidly. Patterns of phosphoproteins in extracts from these labelled parasites were compared after two dimensional electrophoresis and autoradiography. Protein extracts from eggs, microfilariae and adult females of O. volvulus were phosphorylated in the presence of [gamma-32P]ATP, magnesium acetate, and added cyclic AMP-dependent protein kinase or the endogenous protein kinase present in the extracts. Patterns of phosphoproteins were compared after separation by single and two-dimensional gel electrophoresis followed by autoradiography. Common phosphopeptide bands were observed when phosphorylated extracts from adult females, microfilariae and eggs were compared. However, extracts from eggs displayed unique phosphorylated polypeptides of M(r) 30,000 and 34,000 that were absent from the extracts from microfilariae. Furthermore, two phosphorylated polypeptides of M(r) 47,000 and 76,000 were detected in extracts from microfilariae but not from eggs. These results indicate that O. volvulus parasites may phosphorylate different proteins at different stages of their development.

摘要

观察发现,将旋盘尾丝虫的活体雌虫和微丝蚴置于含有[32P]正磷酸盐的培养基中培养时,它们会迅速使自身蛋白质磷酸化。在进行二维电泳和放射自显影后,比较了这些标记寄生虫提取物中的磷蛋白模式。旋盘尾丝虫的卵、微丝蚴和成年雌虫的蛋白质提取物在[γ-32P]ATP、乙酸镁以及添加的环磷酸腺苷依赖性蛋白激酶或提取物中存在的内源性蛋白激酶存在的情况下发生磷酸化。在通过单向和二维凝胶电泳分离并进行放射自显影后,比较了磷蛋白模式。比较成年雌虫、微丝蚴和卵的磷酸化提取物时,观察到了共同的磷酸肽带。然而,卵的提取物显示出微丝蚴提取物中不存在的分子量为30,000和34,000的独特磷酸化多肽。此外,在微丝蚴提取物中检测到了分子量为47,000和76,000的两种磷酸化多肽,但在卵的提取物中未检测到。这些结果表明,旋盘尾丝虫寄生虫在其发育的不同阶段可能会使不同的蛋白质磷酸化。

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