Fine resolution of human sperm nucleoproteins by two-dimensional electrophoresis.
Fine resolution of human sperm nucleoproteins by two-dimensional electrophoresis.
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DOI：
10.1093/molehr/gah217
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发表时间：
2005-09
影响因子：
4
通讯作者：
Tsuyoshi Yoshii;N. Kuji;S. Komatsu;K. Iwahashi;Yudai Tanaka;H. Yoshida;A. Wada;Y. Yoshimura
中科院分区：
文献类型：
--
作者：
Tsuyoshi Yoshii;N. Kuji;S. Komatsu;K. Iwahashi;Yudai Tanaka;H. Yoshida;A. Wada;Y. Yoshimura
人类精子核蛋白由鱼精蛋白和组蛋白组成。这些蛋白质组成的变化被认为与精子发生相关，并且可能与男性不育的某些情况有关。我们试图使用改进的二维电泳方法分离包括鱼精蛋白变体在内的精子核蛋白，目的是全面分析所有精子核蛋白成分。从正常志愿者的精子中提取核碱性蛋白后，我们通过基于卡尔施密特和惠特曼二维电泳的无自由基、高度还原方法在凝胶片上分析这些蛋白质。来自精子核的碱性蛋白质被清楚地分离成 12 个点。通过氨基酸序列分析，这些点对应于鱼精蛋白1（P1）-（5个点）、鱼精蛋白2（P2）相关蛋白（6个点）和睾丸特异性组蛋白H2B（1个点）。 6种P2相关蛋白的N端氨基酸序列与HPI1、HPI2、HPS1、HPS2、HP2和HP3的N端氨基酸序列一致，可以进行定量比较。总之，可以在单一电泳凝胶上以高分辨率对人类精子核蛋白（包括所有 P2 相关变体）进行定量分析。
Human sperm nucleoproteins consist of protamines and histones. Changes in composition of these proteins are thought to correlate with spermatogenesis and may be involved in some instances of male infertility. We sought to separate sperm nucleoproteins including variants of protamine using an improved two-dimensional electrophoretic method, with the aim of comprehensively analysing all sperm nucleoprotein constituents. After extracting nuclear basic proteins from the sperm of normal volunteers, we analysed these proteins on a gel sheet by a radical free, highly reducing method based on Kaltschmidt and Whittmann's two-dimensional electrophoresis. Basic proteins from sperm nuclei were separated clearly into 12 spots. By amino acid sequence analysis, these spots corresponded to protamine 1 (P1)- (five spots), protamine 2 (P2)-related proteins (six spots) and testis-specific histone H2B (one spot). The N-terminal amino acid sequences of the six P2-related proteins were compatible with those of HPI1, HPI2, HPS1, HPS2, HP2 and HP3, and quantitative comparison could be performed. In conclusion, human sperm nucleoproteins including all P2-related variants could be analysed quantitatively with high resolution on a single electrophoretic gel.