Mapping the interaction between oligodendrocyte-enriched G protein-coupled receptor 37 and myelin-associated glycoprotein 公开

Abstract

G protein-coupled receptor 37 (GPR37) is an orphan G protein-coupled receptor (GPCR) that is most highly expressed in myelinating oligodendrocytes. GPR37-knockout mice exhibit increased susceptibility to demyelination and a significant down-regulation of myelin-associated glycoprotein (MAG), a component of myelin, in the brain. We have previously demonstrated that GPR37 and MAG co-immunoprecipitate, indicating a physical interaction between the two proteins. In order to determine which regions of these proteins are necessary to facilitate the physical interaction, we performed a series of co-immunoprecipitation experiments with several truncated variants. These studies revealed that the N-terminal domain of MAG is not required for the interaction, nor the N-terminal and C-terminal domains of GPR37, suggesting that the physical interaction is facilitated by the transmembrane domain regions of each protein. Furthermore, we found that each of the seven individual transmembrane domains of GPR37 co-immunoprecipitates with MAG. This work forms an important basis for understanding the structural determinants of the interaction between GPR37 and MAG and manipulating this interaction for use in future experimental models. Ultimately, GPR37 is a promising potential drug target for demyelinating disorders, and studying its interaction with MAG is important for understanding the role of GPR37 in disease pathology and potential treatments.

Table of Contents

Introduction …………………………………………………………………………………....1

Orphan G protein-coupled receptors as potential drug targets

G protein-coupled receptor 37 (GPR37)

GPR37 in oligodendrocytes

Myelin-associated glycoprotein (MAG)

Structural studies

Figure 1 …………………………………………………………………………….......4

Materials and methods ……………………………………………………………………….5

Cell culture and transfection

Cell lysate preparation

Co-immunoprecipitation studies

Figure 2 ………………………………………………………………………………....6

Western blot analysis

Figure 3 ………………………………………………………………………………….7

Results .………………………………………………………………………………..………....9

Figure 4 ………………………………………………………………………………….9

Figure 5 …………………………………………………………………………...……10

Figure 6 …………………………………………………………………………...……11

Figure 7 …………………………………………………………………………..…….12

Figure 8 ……………………………………………………………………………..….13           

Discussion …………………………………………………………………………………...…14

Figure 9 ………………………………………………………………………………...15

References …………………………………………………………………………………...…20

About this Honors Thesis

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School	Emory College
Department	Biology
Degree	B.S.
Submission	Honors Thesis
Language	English
Research Field	Biology, Neuroscience Biology, Cell Biology, Molecular
关键词	myelin receptor protein-protein interaction GPCR pharmacology
Committee Chair / Thesis Advisor	Randy Hall, Emory University
Committee Members	Roger Deal, Emory University Kristen Frenzel, Emory University

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