Establishment of a high-resolution 2-D reference map of human spermatozoal proteins from 12 fertile sperm-bank donors.
OBJECTIVE
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 3872 different protein spots were identified from the reference map, an approximately 3-fold increase compared to the broad range pH 3-10 IPG strip (1306 spots).


CONCLUSIONS
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.
Li
Ling-Wei
LW
Institute of Reproduction and Stem Cells Engineering, Central-South University, Changsha 410078, China.
Fan
Li-Qing
LQ
Zhu
Wen-Bing
WB
Nien
Hong-Chuan
HC
Sun
Bo-Lan
BL
Luo
Ke-Li
KL
Liao
Ting-Ting
TT
Tang
Le
L
Lu
Guang-Xiu
GX
D000328:Adult; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005298:Fertility / Q000502:physiology; D006801:Humans; D008297:Male; D011506:Proteins / Q000032:analysis*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012016:Reference Values; D012661:Semen / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013074:Sperm Banks; D013094:Spermatozoa / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D014019:Tissue Donors
D011506:Proteins; D020543:Proteome