Structure/Function of the CB2 Receptor Binding and G-protein Recognition Pockets

CB2 受体结合和 G 蛋白识别口袋的结构/功能

• 批准号: 8851758
• 负责人: Xiang-Qun Xie
• 金额: $ 9.6万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8851758
• 关键词: Adenylate Cyclase; Adverse effects; Affinity; Agonist; Algorithms; Anti-Obesity Agents; Appetite Depressants; Autoimmune Diseases; Autoimmune Process; Binding; Binding Sites; Biochemical; Biological Assay; C-terminal; CNR2 gene; Cannabinoids; Cell Signaling Process; Chemicals; Chronic; Complication; Computer Simulation; Coupled; Coupling; Data; Depression and Suicide; Disease; Docking; Drug Design; Drug Targeting; Enzymes; European; Event; Fluorescence; Future; G Protein-Coupled Receptor Genes; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Glioma; Goals; Guanosine Triphosphate Phosphohydrolases; Health; Human; Immune; Immune system; Isotope Labeling; Length; Ligand Binding; Ligands; Light; Marketing; Mediating; Metabolism; Methods; Molecular; Mutagenesis; Mutation; Neurons; Outcomes Research; Pain; Patients; Pharmaceutical Preparations; Pilot Projects; Proteins; Publishing; Recombinants; Reporting; Research; Research Design; Resolution; Roentgen Rays; Signal Transduction; Site; Specificity; Structure; Structure-Activity Relationship; System; Techniques; Therapeutic; Therapeutic Studies; Transmembrane Domain; Validation; Work; base; biophysical techniques; cannabinoid receptor; design; drug discovery; endogenous cannabinoid system; experience; extracellular; interest; novel; pharmacophore; protein expression; receptor; receptor binding; receptor structure function; research study; rimonabant; three dimensional structure; tumor

DESCRIPTION (provided by applicant): The discovery of the endogenous cannabinoid (CB) system, i.e. the CB receptors (the subtypes CB1 and CB2), endogenous ligands and enzymes for CB ligand metabolism, has triggered intensive pharmacological research into the CB receptors and the therapeutic potential of cannabinergic ligands. The CB2 receptor is known to be involved in the signal transduction cascades in the immune system, and has a great therapeutic potential for developing CB2 drugs without CB1-related psychotropic side effects for treatment of chronic neuro-pain, neuronal disorders, autoimmune diseases, and gliomas and other tumors of immune origin, thereby benefiting the millions of patients who suffer from the autoimmune/immune-related diseases for which we have no cure. Over the years, however, structural and functional studies of CB receptors have focused on predicting structures by computer modeling, identification of the specific sites for ligand binding and G-protein coupling whereas studies on experiment 3D CB structures are limited, in particular for the CB2 receptor. To understand the molecular mechanism behind these pharmacological and biochemical events, it is important to characterize the contacting points and binding domains and then elucidate the specific CB2/ligand recognition and CB2/G-protein coupling mechanisms at molecular structural level. In our published and pilot studies, we have successfully investigated the structural and conformational features of several CB2 protein functional domains; CB ligand structures and active pharmacophoric features; and agonist/antagonist recognition sites in the CB2 receptor. However, many questions still remain about CB2 receptor structure-function relationship as well as CB2 ligands and G-protein recognition mechanisms. The objective of this proposal is to identify/characterize the key residues/functional domains and elucidate their 3D structures of recognition pockets important to agonist and antagonist binding as well as G-protein coupling recognitions in the CB2 receptor by the combined biophysical and biochemical approaches. Our long term goal is to understand, in structural and functional terms, the molecular mechanisms of human CB2 activation and G-protein cell signaling process in order to facilitate the structure-based design for novel CB2 ligands. Having completed the "proof-of-concept" research work, we propose the specific goals and in-depth research to three aims. Aim 1: Characterize the functional domains and key residues important to the CB2 ligand recognition and derive the structural determinants of the agonist/antagonist binding domains in the transmembrane and extra-cellular segments. Aim 2: Investigate and define the structural and functional features of the important CB2 intracellular segments and key residues involving G-protein coupling and intracellular cell signaling by the biophysical approaches developed and validated in Aim 1. Aim 3: Explore full-length CB2 receptor and confirm the key residues determined in Aims 1 and 2 and verify their importance to CB2 ligand recognition and G-protein coupling by functional binding assays and site-directed mutations of the native CB2 receptor. The elucidated CB2 agonist/antagonist recognition pockets and G-protein coupling domains will be further examined by our established computer modeling and receptor docking algorithms. Our proposed research and the outcomes will shed light onto a better understanding of CB2 structure/function and its mechanism of actions, and provide the structural bases for CB2-specific drug design in future. The techniques and methods developed from the proposed CB2 receptor research will also have a significant impact to other GPCRs.

描述（由申请人提供）：内源性大麻素（CB）系统的发现，即CB受体（亚型CB1和CB2），内源性配体和CB配体代谢的酶，具有强化的强化药理研究CB受体的药理研究，对CB受体进行了cb受体的潜在和治疗性LIGABINGINBINGANBANBINBINDICEN。已知CB2受体与免疫系统中的信号转导级联有关，并且具有很大的治疗潜力，可以在没有CB1相关的精神副作用的情况下开发CB2药物，用于治疗慢性神经疾病，神经疾病，神经元疾病，自身免疫性疾病和gllioMas和其他因素造成免疫原性的疾病，从而受益于慢性神经疾病，神经元疾病，神经元疾病，自身免疫性的疾病和其他因素。我们无法治愈的自身免疫/免疫相关疾病。然而，多年来，CB受体的结构和功能研究集中在通过计算机建模，鉴定配体结合的特定位点和G蛋白偶联的特定位点的结构和功能研究，而实验3D CB结构的研究尤其是CB2受体的限制。为了了解这些药理和生化事件背后的分子机制，重要的是要表征接触点和结合域，然后在分子结构水平上阐明特定的CB2/配体识别以及CB2/G蛋白偶联机制。在我们已发表的试点研究中，我们成功地研究了几个CB2蛋白功能域的结构和构象特征。 CB配体结构和活跃的药物成分特征；和CB2受体中的激动剂/拮抗剂识别位点。但是，关于CB2受体结构功能关系以及CB2配体和G蛋白识别机制仍然存在许多问题。该提案的目的是识别/表征关键残基/功能域，并阐明其对激动剂和拮抗剂结合至关重要的识别口袋，以及通过合并的生物物理和生物化学方法中CB2受体中CB2受体中的G蛋白偶联识别。我们的长期目标是从结构和功能术语中理解人CB2激活和G蛋白细胞信号传导过程的分子机制，以促进用于新型CB2配体的基于结构的设计。完成了“概念证明”研究工作后，我们提出了三个目标的具体目标和深入研究。目标1：表征对CB2配体识别重要的功能域和关键残基，并得出跨膜和细胞外段中激动剂/拮抗剂结合域的结构决定因素。目标2：研究并定义重要CB2重要的细胞内片段的结构和功能特征，以及涉及G蛋白耦合和细胞内细胞信号传导的关键残基，并通过目标1中开发和验证的生物物理方法。AIM3：AIM 3：探索全长的CB2受体，并确认在AIM 1和2中确定CB2的关键残基，并确定CB2 lig 2 lig2 lig2 lig 2 lig 2天然CB2受体的结合测定和位置定向的突变。我们已建立的计算机建模和受体对接算法将进一步检查阐明的CB2激动剂/拮抗剂识别口袋和G蛋白耦合域。我们提出的研究和结果将使人们对CB2结构/功能及其作用机理有更好的了解，并为将来的CB2特异性药物设计提供结构碱基。拟议的CB2受体研究开发的技术和方法也将对其他GPCR产生重大影响。

项目成果

Structural insight into tetrameric hTRPV1 from homology modeling, molecular docking, molecular dynamics simulation, virtual screening, and bioassay validations.

• DOI: 10.1021/ci5007189
• 发表时间: 2015-03-23
• 期刊: Journal of chemical information and modeling
• 影响因子: 5.6
• 作者: Feng Z;Pearce LV;Xu X;Yang X;Yang P;Blumberg PM;Xie XQ
• 通讯作者: Xie XQ

Metal binding mediated conformational change of XPA protein:a potential cytotoxic mechanism of nickel in the nucleotide excision repair.

• DOI: 10.1007/s00894-016-3017-x
• 发表时间: 2016-07
• 期刊: Journal of molecular modeling
• 影响因子: 2.2
• 作者: Hu J;Hu Z;Zhang Y;Gou X;Mu Y;Wang L;Xie XQ
• 通讯作者: Xie XQ

Computational target fishing: what should chemogenomics researchers expect for the future of in silico drug design and discovery?

• DOI: 10.4155/fmc.14.5
• 发表时间: 2014-03
• 期刊: Future medicinal chemistry
• 影响因子: 4.2
• 作者: Wang L;Xie XQ
• 通讯作者: Xie XQ

Comparative normal/failing rat myocardium cell membrane chromatographic analysis system for screening specific components that counteract doxorubicin-induced heart failure from Acontium carmichaeli.

• DOI: 10.1021/ac500287e
• 发表时间: 2014-05-20
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Chen, Xiaofei;Cao, Yan;Zhang, Hai;Zhu, Zhenyu;Liu, Min;Liu, Haibin;Ding, Xuan;Hong, Zhanying;Li, Wuhong;Lv, Diya;Wang, Lirong;Zhuo, Xianyi;Zhang, Junping;Xie, Xiang-Qun;Chai, Yifeng
• 通讯作者: Chai, Yifeng

Cardiovascular Disease Chemogenomics Knowledgebase-guided Target Identification and Drug Synergy Mechanism Study of an Herbal Formula.

心血管疾病化学基因组学知识库引导的中药方靶点识别及药物协同机制研究

• DOI: 10.1038/srep33963
• 发表时间: 2016-09-28
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Zhang H;Ma S;Feng Z;Wang D;Li C;Cao Y;Chen X;Liu A;Zhu Z;Zhang J;Zhang G;Chai Y;Wang L;Xie XQ
• 通讯作者: Xie XQ