CHARACTERIZATION OF PUTATIVE MAGNESIUM TRANSPORTING P-TYPE ATPASES

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。 前言：p型ATPase包括一个蛋白质超家族，存在于原核生物和真核生物中，可以跨细胞膜运输无机阳离子和可能的其他底物。根据系统发育研究，它们被划分为5个亚家族，称为P1-P5，根据序列相似性、跨膜组织和底物特异性进行分组。最不了解的P型ATPase是P5亚家族的那些，它们只在真核生物中表达。目前这项提案的总体目标是获得关于小鼠P5-ATPase的基本信息，这些信息将对最终了解它们的细胞生物学和生理功能至关重要。本方案的目的是1)分离每个家族成员的全长cDNA克隆，并确定它们的组织分布和膜位置；2)在适当的细胞系中过表达新的P型ATPase，以评估它们的 3)利用RNA干扰(RNAi)和基因打靶技术确定它们在体内的细胞和生理作用。

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. Introduction: P-type ATPases comprise a large superfamily of proteins, present in both prokaryotes and eukaryotes, that transport inorganic cations and possibly other substrates across cell membranes. Based on phylogenetic studies they have been classified into 5 subfamilies, termed P1-P5, which group according to sequence similarities, transmembrane organization, and substrate specificity. The most poorly understood P-type ATPases are those of the P5 subfamily, which are expressed only in eukaryotes. The overall goal of the current proposal is to obtain basic information about the P5-ATPases in mice that will be essential for an eventual understanding of their cell biological and physiological functions. The aims of the current proposal are to 1) isolate full-length cDNA clones for each of the family members and determine their tissue distribution and membrane location, 2) overexpress the novel P-type ATPases in the appropriate cell lines in order to assess their i on specificity, and 3) determine their cellular and physiological roles in vivo using RNA interference (RNAi) and gene targeting technologies.