A kind of G protein-coupled receptor living body tracing method and application

Abstract

The invention relates to the field of molecular Imaging, in particular to a G protein-coupled receptor (GPCR) in-vivo tracking method. The GPCR in-vivo tracking method is characterized by inserting a molecular enhanced green fluorescent protein (eGFP) to a non-functional region of the GPCR to construct a novel GPCR-eGFP fusion protein for imaging and tracking of GPCR molecules. In the GPCR in-vivo tracking method, once mutating, rhodopsin which is a photoreceptor protein existing on retinal rod cell membrane of vertebrate widely may cause failure of protein membrane insertion, misfolding and even aggregation and the like to further cause severe eye diseases; therefore, construction of the rhodopsin-eGFP fusion protein and detection distribution of rhodopsin in cells by means of fluorescence are great significant in research on eye diseases. As the molecular eGFP is inserted to the non-functional region, namely cytoplasmic loop 1 (CL1), of rhodopsin originally, the novel rhodopsin CL1-eGFP fusion protein, which is constructed by the method, is of expression level and function similar to those of wild-type proteins, and is an effective tool for research on eye diseases related to rhodopsin.

Description

technical field

[0001] The present invention relates to the field of molecular imaging, specifically a novel G protein-coupled receptor (G Protein-Coupled Receptor, GPCR) tracing method in vivo. Taking human rhodopsin as an example, it is specifically described in its non-functional region, namely the first A molecule of eGFP was inserted into Cytoplasmic Loop 1 (CL1), and a new type of rhodopsin was successfully constructed CL1 -eGFP fusion protein, the intracellular distribution of rhodopsin is detected by fluorescence. Background technique

[0002] G Protein-Coupled Receptor (GPCR) is an important protein that exists on the cell membrane and participates in cell signal transduction, and participates in a variety of important physiological processes, such as synaptic transmission, cell proliferation and differentiation, and Taste and touch perception etc. GPCRs are composed of seven transmembrane helices, which are connected by three intracellular loops and three extrace...

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