Cannabinoid Receptor Interacting Protein 1a (CRIP1a) in cell signaling and cargo transport

细胞信号传导和货物运输中的大麻素受体相互作用蛋白 1a (CRIP1a)

• 批准号: 10612812
• 负责人: Erin K Hughes
• 金额: $ 4.77万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612812
• 关键词: Address; Adolescence; Age; Agonist; Antibodies; Binding; Binding Proteins; Biochemical; Biochemistry; Biological; Brain region; C-terminal; CNR1 gene; Cannabis; Cell model; Cells; Central Nervous System; Collaborations; Complex; Cytoplasm; Data; Defect; Development; Dissociation; Family member; Fluorescence Polarization; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP Binding; GTP-Binding Proteins; Heterotrimeric G Protein Subunit; Heterotrimeric GTP-Binding Proteins; Homologous Gene; Human; Immunoprecipitation; Intracellular Transport; Knowledge; Legal; Ligands; Lipids; Mediating; Medical Marijuana; Membrane; Molecular; Molecular Conformation; Monomeric GTP-Binding Proteins; Mus; N-terminal; Neurons; Neuropharmacology; Pathway interactions; Peptides; Persons; Process; Proliferating; Protein Farnesylation; Protein Isoprenylation; Protein Subunits; Proteins; Receptor Signaling; Regulation; Research Personnel; Resolution; Role; Safety; Schizophrenia; Self Medication; Signal Transduction; Structure; Testing; Tetrahydrocannabinol; Time; Training; X-Ray Crystallography; antagonist; cannabinoid receptor; cannabinoid receptor interacting protein 1a; career development; child bearing; endogenous cannabinoid system; experimental study; farnesylation; inhibitor; innovation; interest; marijuana legalization; marijuana use; marijuana user; migration; myristoylation; nerve stem cell; neural; neuroblastoma cell; neuron development; neurosensory; neurotransmitter release; novel; palmitoylation; protein activation; protein complex; receptor; receptor function; response; synaptogenesis

PROJECT SUMMARY This new F31 proposal is to support the career development of Erin Hughes who will receive training in cannabinoid receptor biochemical neuropharmacology with Drs. Howlett and Lowther. Recent years have seen a global increase in the usage of marijuana containing the psychoactive constituent Δ9-tetrahydrocannabinol, a partial agonist of the CB1 cannabinoid receptor (CB1R). The CB1R is a G-protein coupled receptor (GPCR) found in the central nervous system and modulates neuroprogenitor development, neural commitment and migration, and neurotransmitter release. GPCR function is dependent upon the ability to change conformations and to transduce a response via G-proteins. This process requires association of a Gα-GDP subunit with Gβγ subunits to form a heterotrimeric G-protein complex that binds to the GPCR. Stimulation of CB1R initiates the exchange of GDP for GTP bound to Gα, activating the G-protein heterotrimer, and releasing the Gα and Gβγ subunits. The released G-protein subunits propagate the signaling cascade through various interactions. Cannabinoid receptor interacting protein 1a (CRIP1a) is a protein found in human neurons at various brain regions, which suppresses CB1R cellular signaling via modulating G-protein activation. The Scientific Premise is that CRIP1a modulation of CB1R can be understood providing us with an in-depth view of the role of CB1R in neuronal cell signaling. CRIP1a can modulate the type of Gα subunit bound to CB1R, however the mechanism of switching is unknown. Previous immunoprecipitation studies demonstrated that the CB1R C-terminal region interacts with CRIP1a, and therefore one possible explanation for CRIP1a-mediated Gα switching may involve formation of a ternary complex between CB1R, subtype-specific Gα proteins, and CRIP1a. The Lowther-Howlett collaboration has determined the high-resolution structure of CRIP1a by X-ray crystallography and found that CRIP1a is a member of the family of carriers for myristoylated or isoprenylated proteins. Based on the knowledge of the structure and function of CRIP1a, I hypothesize that CRIP1a binds N-terminally myristoylated cargo proteins, such as Gαi proteins. I propose to investigate CRIP1a interactions in the N18TG2 neuroblastoma cell model which endogenously expresses the CB1R and associated proteins. The aims of this project are to determine (i) the regulation of the CRIP1a- Gα interaction during agonist stimulation and heterotrimeric G protein dissociation and the cellular localization of such interactions; and (ii) the role of palmitoylation and myristoylation on G-protein interaction with CRIP1a. The results of the proposed studies will prove to be transformative for the field by characterizing a new class of cargo carrying proteins and providing evidence that will advance our understanding of how, when and where CRIP1a interactions modulate CB1R pathways.

项目摘要 这个新的F31提案是为了支持艾琳休斯的职业发展，他将接受培训， 大麻素受体生化神经药理学与豪利特和洛瑟博士。近年来 全球使用含有精神活性成分Δ9-四氢大麻酚的大麻的人数增加， CB 1大麻素受体（CB 1 R）的部分激动剂。CB 1 R是一种G蛋白偶联受体（GPCR）， 在中枢神经系统中并调节神经祖细胞发育、神经定型和迁移， 和神经递质的释放GPCR的功能取决于改变构象的能力， 通过G蛋白的免疫应答。这一过程需要Gα-GDP亚基与Gβγ亚基的结合 以形成与GPCR结合的异源三聚体G蛋白复合物。刺激CB 1 R启动交换 GTP与Gα结合，激活G蛋白异源三聚体，释放Gα和Gβγ亚基。的 释放的G蛋白亚基通过各种相互作用传播信号级联。大麻素受体 相互作用蛋白1a（CRIP 1a）是在人类神经元中发现的蛋白质，其抑制 CB 1 R通过调节G蛋白激活的细胞信号传导。科学假设是CRIP 1a调节 CB 1 R在神经元细胞中的作用，为我们提供了一个深入的了解 信号CRIP 1a可调节与CB 1 R结合的Gα亚基的类型，但其转换机制尚不清楚。 不明先前的免疫沉淀研究表明，CB 1 R C-末端区域与 因此，CRIP 1a介导的Gα转换的一种可能解释可能涉及CRIP 1a介导的Gα转换的形成。 CB 1 R、亚型特异性Gα蛋白和CRIP 1a之间的三元复合物。Lowther-Howlett合作 通过X射线晶体学确定了CRIP 1a的高分辨率结构，并发现CRIP 1a是一种 肉豆蔻酰化或异戊二烯化蛋白质载体家族成员。基于对 CRIP 1a的结构和功能，我假设CRIP 1a结合N-末端豆蔻酰化货物 蛋白质，如Gαi蛋白质。我建议研究CRIP 1a在N18 TG 2神经母细胞瘤中的相互作用 内源性表达CB 1 R和相关蛋白的细胞模型。该项目的目的是 确定（i）在激动剂刺激和异源三聚体期间CRIP 1a-G α相互作用的调节 G蛋白解离和这种相互作用的细胞定位;和（ii）棕榈酰化的作用 以及豆蔻酰化对G蛋白与CRIP 1a相互作用的影响。拟议研究的结果将证明， 通过表征一类新的携带蛋白质的货物并提供证据， 这将促进我们对CRIP 1a相互作用如何、何时以及在何处调节CB 1 R通路的理解。