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Phosphorylation-independent Desensitization of mGluR5 Mediated by GRK2

Author: DingHaiLi
Tutor: LiuJianFeng
School: Huazhong University of Science and Technology
Course: Biochemistry and Molecular Biology
Keywords: G protein-coupled receptor kinase 2 G protein -coupled receptors Metabotropic glutamate receptor 5 Phosphorylation Desensitization
CLC: Q42
Type: Master's thesis
Year: 2007
Downloads: 42
Quote: 0
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Abstract


passed for the formation of excitatory nerve mGluRs in glutamatergic synapses play an important role , and mGluRs desensitization is an important mechanism for regulating the function of these receptors . GRKs have confirmed the regulatory role in a variety of GPCRs desensitization from . GRKs and second messenger -dependent protein kinase -mediated mGluR1 desensitization of GRK2 mediated mGluR1 desensitization is not dependent on phosphorylation and β -arrestin . These findings suggest a novel mechanism for an the GRK2 adjustment mGluR5 receptor function in neurotransmission regulation is of great significance . In this issue , we GRK2 and mGluR5 as the research object , GRK2 -mediated mGluR5 signal conditioning were discussed . Molecular cloning techniques to build the missing kinase activity of GRK2 mutant GRK2-K220R and the lack of phosphorylation sites mGluR5 mutant mGluR5-del847-T840A, mGluR5/mGluR5-del847-T840A individually transfected HEK293 cells were detected by ELISA the expression and the expression amount of the co-transfected with the GRK2/GRK2-K220R , and then use the IP3 assay receptor signaling activity is generated , the results show GRK2 mediated the mGluR5 unphosphorylated dependent desensitization , and this loss of sensitization effect is not due to changes in the amount of receptor expression . Meanwhile, we also constructed with a full-length HA antigen markers and 5 contain of GRK2 different functional domains shear by ELISA and Western Blot to successfully expressed in HEK293 cells for subsequent detection which part of GRK2 domains are involved GRK2 -mediated receptor desensitization provides a foundation . Overall , our study confirms the non-phosphorylated dependent GRK2 -mediated mGluR5 desensitization , GRK2 on the receptor desensitization regulation not GRK2 affect the amount of receptor expression , caused these results for subsequent in-depth research GRK2 -mediated GPCRs desensitization mechanism laid the foundation.

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