CONFIRMATION OF SP56 PROTEIN IN HUMAN SPERM

人类精子中 SP56 蛋白的确认

• 批准号: 8171368
• 负责人: WILLIE J SWANSON
• 金额: $ 0.14万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171368
• 关键词: Cell surface; Computer Retrieval of Information on Scientific Projects Database; Databases; Funding; Furuncles; Genes; Grant; Human; Institution; Mus; Orthologous Gene; Proteins; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Sampling; Seminal fluid; Source; Translating; United States National Institutes of Health; egg; sperm cell; sperm protein; spermadhesin; zona pellucida glycoprotein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. sp56 is a rodent specfic sperm protein that binds to ZP3 on the egg cell surface in mice. We have predicted a human ortholog of sp56 that is located in the same syntenic block as mouse sp56. We have confirmed that human sp56 is expressed by RT-PCR and now want to confirm that the translated protein is present in human sperm by mass spec. To prepare the samples, we pelleted sperm from human seminal fluid and solubilized the sperm proteins by boiling them in either SDS or RapiGest. We searched the resulting spectra against the human database containing our predicted sp56 gene to confirm protein presence.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 sp56 是一种啮齿动物特异性精子蛋白，可与小鼠卵细胞表面的 ZP3 结合。我们预测了 sp56 的人类直系同源物，它与小鼠 sp56 位于同一同线区。我们已经通过 RT-PCR 确认了人类 sp56 的表达，现在想要通过质谱确认翻译的蛋白质存在于人类精子中。为了制备样品，我们从人类精液中沉淀出精子，并通过在 SDS 或 RapiGest 中煮沸来溶解精子蛋白。我们根据包含我们预测的 sp56 基因的人类数据库搜索了所得光谱，以确认蛋白质的存在。