Establishment of a high-resolution 2-D reference map of human spermatozoal proteins from 12 fertile sperm-bank donors |
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Authors: | Li Ling-Wei Fan Li-Qing Zhu Wen-Bing Nien Hong-Chuan Sun Bo-Lan Luo Ke-Li Liao Ting-Ting Tang Le Lu Guang-Xiu |
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Affiliation: | Institute of Reproduction and Stem Cells Engineering, Central-South University, Changsha 410078, China. |
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Abstract: | 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 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). 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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Keywords: | human spermatozoal proteins ImageMaster mass spectrometry overlapping narrow range 2-D gel electrophoresis |
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