Establishment of a high-resolution 2-D reference map of human spermatozoal proteins from 12 fertile sperm-bank donors
Establishment of a high-resolution 2-D reference map of human spermatozoal proteins from 12 fertile sperm-bank donors作者机构：Institute of Reproduction and Stem Cells Engineering Central-South University Changsha 410078 China
出 版 物：《Asian Journal of Andrology》 (亚洲男性学杂志（英文版）)
年 卷 期：2007年第9卷第3期
页 面：321-329页
核心收录：
学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 1001[医学-基础医学(可授医学、理学学位)] 07[理学] 08[工学] 0817[工学-化学工程与技术] 10[医学]
基 金：This work was supported by two grants from the National Natural Science Foundation of China (No. 30170480 and No. 30470884). We would like to thank Professor Guang-Yin Lu for enthusiastic reading and amending of the manuscript. We would like to thank Mr Ji-Xian Xiong College of Life Science at the Hunan Normal University for enthusiastic support of technology and theory of MS. We are also very grateful to collaborators in our laboratories for help and valuable discussions and suggestions during the course of this work
主 题：human spermatozoal proteins ImageMaster mass spectrometry overlapping narrow range 2-D gel electrophoresis
摘 要：Aim： To extend the analysis of the proteome of human spermatozoa and establish a 2-D gel electrophoresis （2-DE） reference map of human spermatozoal proteins in a pH range of 3.5-9.0. Methods： In order to reveal more protein spots, immobilized pH gradient strips （24 cm） of broad range of pH 3-10 and the narrower range of pH 6-9, as well as different overlapping narrow range pH immobilized pH gradient （IPG） strips, including 3.5-4.5, 4.0-5.0, 4.5-5.5, 5.0-6.0 and 5.5-6.7, were used. After 2-DE, several visually identical spots between the different pH range 2-D gel pairs were cut from the gels and confirmed by mass spectrometry and used as landmarks for computer analysis. Results： The 2-D reference map with pH value from 3.5 to 9.0 was synthesized by using the ImageMaster analysis software. The overlapping spots were excluded, so that every spot was counted only once. A total of 3 872 different protein spots were identified from the reference map, an approximately 3-fold increase compared to the broad range pH 3-10 IPG strip （1 306 spots）. Conclusion： The present 2-D pattern is a high resolution 2-D reference map for human fertile spermatozoal protein spots. A comprehensive knowledge of the protein composition of human spermatozoa is very meaningful in studying dysregulation of male fertility.
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