Open AccessEstablishment of a high‐resolution 2‐D reference map of human spermatozoal proteins from 12 fertile sperm‐bank donors
Author(s) -
Li LingWei,
Fan LiQing,
Zhu WenBing,
Nie HongChuan,
Sun BoLan,
Luo KeLi,
Liao TingTing,
Tang Le,
Lu GuangXiu
Publication year - 2007
Publication title -
asian journal of andrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.701
H-Index - 74
eISSN - 1745-7262
pISSN - 1008-682X
DOI - 10.1111/j.1745-7262.2007.00261.x
Subject(s) - spots , immobilized ph gradient , resolution (logic) , proteome , electrophoresis , biology , sperm , chemistry , chromatography , microbiology and biotechnology , biochemistry , enzyme , genetics , artificial intelligence , isoelectric focusing , computer science , botany
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.