The rat sperm proteome characterized via IPG strip prefractionation and LC-MS/MS identification
The rat sperm proteome characterized via IPG strip prefractionation and LC-MS/MS identification
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DOI：
10.1002/pmic.200700876
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发表时间：
2008-06-01
期刊：
影响因子：
3.4
通讯作者：
Aitken, R. John
中科院分区：
文献类型：
--
作者：
Baker, Mark A.;Hetherington, Louise;Aitken, R. John
蛋白质组学是一个强大的工具，哺乳动物精子的分析，因为这些终末分化的细胞是转录失活，并表现出有限的动态范围的蛋白质表达。在这里，我们报告的5123肽的鉴定，导致829明确和2215冗余的基因产物被发现存在于大鼠附睾尾精子。生物信息学表明，60种蛋白质似乎在泌尿生殖道中特异性表达，包括丙酮酸脱氢酶1、ropporin、睾丸特异性丝氨酸激酶4、睾丸特异性转运蛋白和视黄醇脱氢酶14。我们还鉴定了ADAM家族的8个成员，其中7个先前在精子中检测到（ADAM 2、-3、-4、-5、-6、-7和-30），而ADAM 34首次在精子蛋白质组中被鉴定。发现大约21种基因产物具有异构酶活性，包括已知参与生殖细胞分化的肽基脯氨酰顺/反异构酶和涉及精子-卵母细胞融合的蛋白质二硫键异构酶。此外，51个基因产物聚集成离子转运活性。这种基因产物的库存，首次对大鼠精子进行2-D LC-MS/MS分析，将在指导未来研究驱动这些高度特化细胞的分子机制方面具有非常宝贵的价值。
Proteomics represents a powerful tool for the analysis of mammalian spermatozoa, since these terminally differentiated cells are transcriptionally inactive and exhibit a limited dynamic range of protein expression. Here we report the identification of 5123 peptides, leading to 829 unambiguous and 2215 redundant gene products found to be present within rat spermatozoa derived from the cauda epididymis. Bioinformatics demonstrated that 60 proteins appeared to be specifically expressed in the genitourinary tract, including pyruvate dehydrogenase 1, ropporin, testisspecific serine kinase 4, testis-specific transporter, and retinol dehydrogenase 14. We also identified eight members of the ADAM family, seven of which have previously been detected in spermatozoa (ADAM2, -3, -4, -5, -6, -7, and -30) while ADAM34 has been identified in the sperm proteome for the first time. Approximately 21 gene products were found to possess isomerase activity including peptidylprolyl cis/trans isomerases that are known to be involved in germ cell differentiation and protein disulfide isomerases that have been implicated in sperm-oocyte fusion. Furthermore, 51 gene products clustered into ion-transporter activity. This inventory of gene products, the first ever 2-D LC-MS/MS analysis of rat spermatozoa, will be invaluable in directing future research into the molecular mechanisms that drive these highly specialized cells.