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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novel strategies oligodendrocyte precursor postnatal precursor cell receptor receptor binding response success tool unpublished works white matter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.

摘要：细胞与周围的细胞外基质(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。黏附G蛋白家族- 偶联受体(AGPCRs)，是第二大类GPCRs，共有33个成员人类基因组，使细胞能够感觉到它们的基质环境。到目前为止，大多数aGPCR是孤儿受体和为数不多的去孤儿的aGPCRs中的每一个都有多个配体来介导细胞-细胞和细胞外基质互动。AGPCRGPR56/ADGRG1是脑发育和出生后脑连接所必需的 (髓鞘形成和突触形成/细化)。GPR56基因突变导致毁灭性的人脑以皮质发育不良和白质髓鞘减退为特征的畸形。与其他类似 AGPCRs与已知的配体，GPR56有多个结合伙伴，胶原III在发育中的大脑黑色素瘤中的皮质、组织转谷氨酰胺酶(TG2)和培养细胞中的糖胺多糖肝素。体外培养。通过无偏倚的体外生物素化/蛋白质组学方法，我们未发表的工作证明 TG2是少突胶质前体细胞中GPR56的配体。GPR56在多个细胞中表达发育中的大脑中的类型，包括放射状胶质细胞、神经前体细胞、星形胶质细胞、OPC和小胶质细胞。我们假设GPR56通过以下途径在不同细胞类型中执行不同的发育任务响应不同配体的差异信号来调节这些效应。为了开始检验这一假设，我们建议生成基因工具包，使我们最终能够在体内进行生物素化蛋白质组学展现GPR56“互动组”景观。