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On-line Access: 2012-07-05

Received: 2011-11-26

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Crosschecked: 2012-06-05

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Journal of Zhejiang University SCIENCE B 2012 Vol.13 No.7 P.555-566


Sphingosine 1-phosphate acts as an activator for the porcine Gpr3 of constitutively active G protein-coupled receptors

Author(s):  Bao-le Zhang, Ye Li, Jian-hua Ding, Fu-lu Dong, Yan-jun Hou, Bao-chun Jiang, Fang-xiong Shi, Yin-xue Xu

Affiliation(s):  College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

Corresponding email(s):   xuyinxue@njau.edu.cn

Key Words:  G protein-coupled receptor, Constitutive activity, Sphingosine 1-phosphate, Receptor internalization, Porcine Gpr3, Molecular cloning

Bao-le Zhang, Ye Li, Jian-hua Ding, Fu-lu Dong, Yan-jun Hou, Bao-chun Jiang, Fang-xiong Shi, Yin-xue Xu. Sphingosine 1-phosphate acts as an activator for the porcine Gpr3 of constitutively active G protein-coupled receptors[J]. Journal of Zhejiang University Science B, 2012, 13(7): 555-566.

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author="Bao-le Zhang, Ye Li, Jian-hua Ding, Fu-lu Dong, Yan-jun Hou, Bao-chun Jiang, Fang-xiong Shi, Yin-xue Xu",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Sphingosine 1-phosphate acts as an activator for the porcine Gpr3 of constitutively active G protein-coupled receptors
%A Bao-le Zhang
%A Ye Li
%A Jian-hua Ding
%A Fu-lu Dong
%A Yan-jun Hou
%A Bao-chun Jiang
%A Fang-xiong Shi
%A Yin-xue Xu
%J Journal of Zhejiang University SCIENCE B
%V 13
%N 7
%P 555-566
%@ 1673-1581
%D 2012
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1100353

T1 - Sphingosine 1-phosphate acts as an activator for the porcine Gpr3 of constitutively active G protein-coupled receptors
A1 - Bao-le Zhang
A1 - Ye Li
A1 - Jian-hua Ding
A1 - Fu-lu Dong
A1 - Yan-jun Hou
A1 - Bao-chun Jiang
A1 - Fang-xiong Shi
A1 - Yin-xue Xu
J0 - Journal of Zhejiang University Science B
VL - 13
IS - 7
SP - 555
EP - 566
%@ 1673-1581
Y1 - 2012
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1100353

We cloned the complete coding sequences of porcine Gpr3, Gpr6, and Gpr12 genes. Further, on the basis of their high levels of sequence similarity, these genes are identified as a subfamily of g protein-coupled receptors. These putative protein sequences also showed high sequence identity with other mammalian orthologs, including several highly conserved motifs. A wide expression of the Gpr3 gene in pigs was observed through tissue distribution analysis by reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR, specially in the brain, pituitary, fat, liver and oocyte, where its strong expression was observed. The Gpr3 gene was found to be located on chromosome 6 and a single exon coded for the entire open-reading frame. Expression of porcine Gpr3 in HEK293 cells resulted in constitutive activation of adenylate cyclase (AC) similar in amplitude to that produced by fully stimulated Gs-coupled receptors. Moreover, sphingosine 1-phosphate (S1P) could increase AC activation via the constitutively active Gpr3 receptor. When a Gpr3-green fluorescent protein (GFP) construct was expressed in HEK293 cells, GFP-labeled Gpr3 protein was shown to be localized in the plasmalemma and subcellular membranes. After S1P treatment, agonist-mediated internalization could be visualized by confocal microscopy. In short, our findings suggest the porcine Gpr3, Gpr6, and Gpr12 genes as a subfamily of g protein-coupled receptors, and porcine Gpr3 was a constitutively active g protein-coupled receptor. Constitutive activation of AC and agonist-mediated internalization of Gpr3 receptor could be modulated by the S1P, suggesting that S1P might act as an activator for porcine Gpr3 receptor.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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