CONFIRMATION OF SP56 PROTEIN IN HUMAN SPERM

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 SP56是一种啮齿动物特异的精子蛋白，与小鼠卵细胞表面的ZP3结合。我们已经预测了人类sp56的直系同源基因，它与老鼠sp56位于同一共线区块。我们已经通过RT-PCR证实了人sp56的表达，现在想要通过质谱仪确认翻译的蛋白存在于人类精子中。为了准备样品，我们从人类精液中制粒精子，并通过在十二烷基硫酸钠或RapiGest中沸腾来溶解精子蛋白。我们在包含我们预测的sp56基因的人体数据库中搜索得到的光谱，以确认蛋白质的存在。

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.