使用酸性凝胶电泳法进行鱼精蛋白提取及人类精子鱼精蛋白1/鱼精蛋白2比例分析。

Protamine extraction and analysis of human sperm protamine 1/protamine 2 ratio using Acid gel electrophoresis.

作者信息

Liu Lihua, Aston Kenneth I, Carrell Douglas T

机构信息

Andrology and IVF Laboratories, Division of Urology, Department of Surgery, University of Utah School of Medicine, Salt Lake City, UT, USA.

出版信息

Methods Mol Biol. 2013;927:445-50. doi: 10.1007/978-1-62703-038-0_38.

DOI:10.1007/978-1-62703-038-0_38

PMID:22992935

Abstract

Protamines, sperm-specific nuclear proteins, are essential for sperm chromatin condensation and DNA stabilization. They are small, highly basic, and rich in disulfide bonds. Under reducing conditions, protamines, along with other basic proteins, are soluble in acid solutions. Because of their small and similar molecular weights, SDS-PAGE cannot resolve protamine 1 and protamine 2 well. Urea-acid gel electrophoresis separates proteins based on the level of the positive charge and is thus a suitable method for resolving protamines 1 and 2. Here, we describe the commonly used protamine extraction method and the Urea-acid gel electrophoresis for assessment of protamine 1/protamine 2 ratio.

摘要

鱼精蛋白是精子特异性核蛋白，对精子染色质凝聚和DNA稳定至关重要。它们体积小、碱性强且富含二硫键。在还原条件下，鱼精蛋白与其他碱性蛋白一起可溶于酸性溶液。由于它们分子量小且相似，SDS-PAGE无法很好地分离鱼精蛋白1和鱼精蛋白2。尿素-酸性凝胶电泳根据正电荷水平分离蛋白质，因此是分离鱼精蛋白1和2的合适方法。在此，我们描述了常用的鱼精蛋白提取方法以及用于评估鱼精蛋白1/鱼精蛋白2比例的尿素-酸性凝胶电泳。