TOWARDS SOLID-STATE NMR STRUCTURES OF GPCRS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH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 中心，不一定是研究者的机构。 GPCR是在几乎所有人体组织中表达的膜蛋白，并且响应于不同的刺激（包括光、激素、损伤和炎症）而传递各种各样的信号。 这些信号通过与GTP结合蛋白的相互作用调节多种细胞反应。许多急性和慢性疾病状态与GPCR功能或故障有关，40-60%的市售药物与GPCR相互作用，使其成为全球近30%的药物发现工作的目标。 1995年，这些药物的市场总额为840亿美元。到目前为止，已经分离出超过300种GPCR，并且大约150种的功能是已知的。据估计，人类基因组中存在约2000个GPCR。努力了解GPCR的功能，结构，稳定性和装配受到阻碍的困难与生产这些整合膜蛋白。 该COBRE小型项目的目标是：1）建立制备同位素富集的人A2 a和人NK 1受体的条件，使其处于功能活性形式，并适合于通过NMR光谱进行结构分析，以及2）获得上述受体的初步多维溶液或固态NMR光谱，用于随后的结构分析。 这些努力的长期目标是获得这些受体的原子级结构信息。 该项目将加速膜蛋白GPCR家族的结构研究，并使人们能够更好地理解这类迷人的蛋白质的配体识别。

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. GPCRs are membrane proteins expressed in virtually all human tissues, and transmit a wide variety of signals in response to diverse stimuli (including light, hormones, injury, and inflammation). These signals regulate a diverse set of cellular responses via interaction with GTP-binding proteins. Many acute and chronic disease states are linked to GPCR function or malfunction, and 40-60% of commercially available drugs interact with a GPCR, making them targets for nearly 30% of drug discovery efforts worldwide. Together these drugs had an $84 billion market in 1995. Thus far over 300 GPCRs have been isolated, and the functions of about 150 are known. It is estimated that ~2000 GPCRs exist in the human genome. Efforts to understand GPCR function, structure, stability, and assembly are hampered by the difficulties associated with producing these integral membrane proteins. The goals of this COBRE mini-project are: 1) to establish the conditions for preparing isotopically enriched human A2a and human NK1 receptors in their functionally competent forms, and suitable for structural analysis by NMR spectroscopy, and 2) to acquire preliminary multidimensional solution or solid-state NMR spectra of the above receptors, for subsequent structural analysis. The long term goal of these efforts is to gain atomic-level structural information of these receptors. This project will accelerate the structural studies of the GPCR family of membrane proteins and enable an improved understanding of the ligand discrimination of this fascinating class of proteins.