LIGAND-RECEPTOR SYSTEMS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们已经开始检查用于流式细胞术的珠上展示的受体和信号分子（Nolan和Sklar，1998; Sklar等人2000）。我们已经展示了His标记的受体可以与Ni 2+珠结合并结合配体的条件。我们还表明，可溶性受体可以用异源三聚体G蛋白（来自Neubig的亚基）和抑制蛋白（Prossnitz）重建，用于荧光，并可能用于流式细胞术分析。 我们已经尝试使用抑制蛋白、受体以及生物素化的受体尾肽作为珠上的诱饵。我们正在检查生物素化的抗his抗体，而不是螯合的Ni 2+珠。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have begun to examine both receptors and signaling molecules displayed on beads for flow cytometry (Nolan and Sklar, 1998; Sklar et al. 2000). We have shown conditions under which his-tagged receptors may be bound to Ni2+ beads and bind ligand. We have also shown that solublized receptors may be reconstituted with heterotrimeric G proteins (subunits from Neubig) and arrestins (Prossnitz) for fluorescence, and potentially for flow cytometric analysis. We have attempted to use arrestin, receptors, as well as biotinylated receptor tail peptides as bait on beads. We are examining biotinylated anti-his antibodies instead of chelated Ni2+ beads.