[Subcellular localization and tissues expression profile of hGPCRc: an orphan G protein-coupled receptor].
As a member of orphan G protein-coupled receptors (oGPCRs), hGPCRc was cloned from human colon tissue and analyzed by bioinformatic softwares. It was showed that the corresponding amino acids of hGPCRc formed seven-transmembrane domains as the key characteristic of GPCRs. Then, the recombinant GFP-hGPCRc was constructed by fussing hGPCRc into pEGFP-N1 carrying green fluorescent protein (GFP) gene, and CHO-K1 cells were subsequently transfected with the GFP-hGPCRc or pEGFP-N1. The green fluorescence protein expression in the two different transfected cells was observed under the laser scanning confocal microscopy (LSCM). It was showed that green fluorescence protein was distributed in the whole bodies of the cells transfected with pEGFP-N1, but mainly distributed on the plasma membrane and cytoplasm membrane transfected with GFP-hGPCRc. Thus, the localization on the membrane of hGPCRc was accorded with the predication by bioinformatic analysis. The expression analysis of hGPCRc by RT-PCR indicated that hGPCRc was abundantly expressed in heart, kidney, cerebel and colon etc., but absent in liver, cerebra, small intestine and muscle etc. The expressing profile of hGPCRc could provide some useful clues to understanding its effects on embryonic development and physiological functions.
Yuan
Guang-Sheng
GS
Institute of Maize, Sichuan Agricultural University, Ya'an 625014, China.
Pan
Guang-Tang
GT
Wu
Fang-Ming
FM
Han
Chun-Guang
CG
Huang
Huo-Gao
HG
Hu
Ming
M
Sheng
Li
L
Chen
Jing
J
Liu
Yong-Xue
YX
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D014018:Tissue Distribution; D014162:Transfection
C531500:GPR22 protein, human; D043562:Receptors, G-Protein-Coupled; D049452:Green Fluorescent Proteins