LONG TIME-SCALE DYNAMICS OF G-PROTEIN COUPLED RECEPTORS IN EXPLICIT LIPID ENVIR

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 G蛋白偶联受体（GPCR）是细胞与外界环境进行通讯的最大的膜蛋白家族。缺乏良好的表达水平和结晶问题阻碍了获得这些蛋白质的实验结构，这些蛋白质构成了目前药物靶标的约50%。我们的小组已经开发了方法[1]来获得这些GPCR在载脂蛋白和配体结合形式的结构。为了获得对这些蛋白质功能的详细分子视角，在不存在和存在配体的情况下，在其天然细胞环境中以激活和失活形式采样和理解其结构是至关重要的。已知与激活相关的构象变化发生在这些受体中的时间尺度为微米至毫秒，并且仍然超出当前全原子分子动力学模拟的范围。开发资助申请将侧重于使用NAMD [2]进行这些模拟，以实验验证这些蛋白质的结构模型所需的时间尺度，并测试载脂蛋白和配体结合蛋白在显式脂质环境中的稳定性。这些蛋白质的构象变化也将研究使用转向动力学，探索其激活和失活形式之间的相互转换。从该开发补助金中获得的信息将帮助我们开发一个完整的MRAC/LRAC提案，重点关注一些药物靶GPCR，我们已经获得了载脂蛋白和配体结合形式的结构，并且对动力学和功能的理解将提供潜在的候选药物线索，可以作为选择性和有效的激动剂或拮抗剂。[1]Vaidehi等人，Proc.Natl. Acad. Sci. USA 99，12622（2002）[2]菲利普斯等人J. Comp. Chem. 26，1781（2005）。

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. G-protein coupled receptors (GPCRs) constitute the largest family of membrane proteins used by cells to communicate with their environment. Lack of good expression levels and crystallization issues have obstructed obtaining experimental structures for these proteins, that constitute about 50% of the current pharmaceutical targets. Our group has developed methods [1] to obtain structures of these GPCRs in apo and ligand-bound forms. To gain a detailed molecular perspective on the function of these proteins, it is critical to sample and understand their structures in activated and deactivated forms in their native cell environment, both in the absence and presence of ligands. Conformational changes, which are associated with activation, are known to occur in these receptors on micro to millisecond time scales and are still beyond the reach of current all-atom molecular dynamics simulations. This application for a development grant will focus on performing these simulations using NAMD [2] to experiment with the time scales necessary and sufficient to validate structural models of these proteins and to test the stability of apo and ligand-bound proteins in explicit lipid environment. The conformational changes in these proteins will also be studied using steered dynamics to explore interconversions between their activated and inactivated forms. The information obtained from this development grant will assist us in developing a full MRAC/LRAC proposal to focus on a few pharmaceutical target GPCRs, for which we have obtained structures in apo and ligand-bound forms, and for which an understanding of dynamics and function will provide potential drug candidate leads that can act as selective and effective agonists or antagonists. [1] Vaidehi et al. Proc. Natl. Acad. Sci. USA 99, 12622 (2002) [2] Phillips et al. J. Comp. Chem. 26, 1781 (2005).