Adhesion GPCR interactome landscaping by in vivo biotinylation proteomics

通过体内生物素化蛋白质组学进行粘附 GPCR 相互作用组景观美化

基本信息

批准号：
9750292
负责人：
Xianhua Piao
金额：
$ 20.07万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9750292
关键词：
ADGR1 gene Address Adhesions Amino Acids Antigen-Antibody Complex Astrocytes Behavior Binding Biological Assay Biology Biotin Biotinylation Birth Rate Brain Cancer Biology Cardiovascular system Cell physiology Cells Cerebral cortex Collagen Communities Cortical Dysplasia Cultured Cells Cytosol Development Developmental Process Disease Embryo Endocrinology Ensure Environment Epitopes Extracellular Matrix Family Family member G-Protein-Coupled Receptors Genetic Genomics Glycosaminoglycans Hematopoiesis Heparin Human Human Genome Immunology Immunoprecipitation In Vitro Joints Ligands Ligase Malignant Neoplasms Mass Spectrum Analysis Mediating Methods Microglia Modeling Morphology Mouse Strains Mus Mutation Neocortex Neuroglia Neurosciences Orphan Pathway interactions Peptide Signal Sequences Phenotype Proteins Proteomics Radial Research Signal Transduction Site Stains Streptavidin Synapses Testing Tissues Transgenic Mice Transglutaminases Weight base body system brain malformation brain morphology cell motility cell type extracellular in vivo innovative technologies melanoma member myelination nerve stem cell novel strategies oligodendrocyte precursor postnatal precursor cell receptor receptor binding response success tool unpublished works white matter

项目摘要

ABSTRACT: The interactions between cells and their surrounding extracellular matrix (ECM) regulate various cellular processes, including morphology, differentiation, motility, and fate. The family of adhesion G protein- coupled receptors (aGPCRs), comprising the second largest class of GPCRs with a total of 33 members in the human genome, enables cells to sense their matrix environment. To date, majority of aGPCRs are orphan receptors and each of the few de-orphaned aGPCRs have multiple ligands to mediate cell-cell and ECM interactions. The aGPCR GPR56/ADGRG1 is essential for brain development and postnatal brain wiring (myelination and synaptic formation/refinement). Mutations in GPR56 cause a devastating human brain malformation that is characterized by cortical dysplasia and white matter hypomyelination. Similar to other aGPCRs with known ligands, GPR56 has multiple binding partners, collagen III in the developing cerebral cortex, tissue transglutaminase (TG2) in melanoma, and glycosaminoglycan heparin in cultured cells in vitro. Through unbiased in vitro biotinylation/proteomics approach, our unpublished work demonstrated that TG2 is the ligand of GPR56 in oligodendrocyte precursor cells (OPCs). GPR56 is expressed in multiple cell types in the developing brain, including radial glial cells, neural progenitor cells, astrocytes, OPCs, and microglia. We hypothesize that GPR56 carries out distinct developmental tasks in different cell types via differential signaling in response to varied ligands to mediate these effects. To begin to test this hypothesis, we propose to generate genetic tool kits that will allow us to eventually carry out in vivo biotinylation proteomics to reveal GPR56 “interactome” landscape.

抽象的：细胞及其周围细胞外基质（ECM）之间的相互作用调节各种细胞过程，包括形态，分化，运动和命运。粘合剂G蛋白的家族耦合受体（AGPCR），完成了第二大GPCR，总共有33位成员人基因组，使细胞能够感知其基质环境。迄今为止，大多数AGPCR是孤儿受体和少数去甲形的AGPCR中的每一个都有多种配体用于介导细胞细胞和ECM 互动。 AGPCR GPR56/ADGRG1对于大脑发育和产后脑接线至关重要（髓鞘形成和突触形成/精致）。 GPR56中的突变导致毁灭性的人脑以皮质发育不良和白质性降低为特征的畸形。类似于其他 GPR56具有已知配体的AGPCR，具有多个结合伴侣，在发育中的大脑中有胶原蛋白III 皮质，黑色素瘤中的组织转谷氨酰胺酶（TG2）和培养细胞中的糖胺聚糖肝素体外。通过公正的体外生物素化/蛋白质组学方法，我们未发表的工作表明 TG2是少突胶质细胞前体细胞（OPC）中GPR56的配体。 GPR56在多个单元中表达发育中的大脑中的类型，包括径向神经胶质细胞，神经祖细胞，星形胶质细胞，OPC和小胶质细胞。我们假设GPR56通过响应各种配体的差异信号传导介导这些作用。为了开始检验这一假设，我们提出生成遗传工具套件的建议，这将使我们最终执行体内生物素化蛋白质组学揭示GPR56“ Interactome”景观。