The C-terminal Phosphorylation Sites of eel Follicle-Stimulating Hormone Receptor are Important Role in the Signal Transduction

Balsaeng'gwa saengsig
Jeong-Min KimKwan-Sik Min

Abstract

The large extracellular domain of glycoprotein hormone receptors is a unique feature within the G protein-coupled receptors (GPCRs) family. After interaction with the hormone, the receptor becomes coupled to Gs, which, in turn stimulates adenylyl cyclase and the production of cAMP. Potential phosphorylation sites exist in the C-terminal region of GPCRs. The experiments described herein represent attempts to determine the functions of the eel follicle-stimulating hormone receptor (eelFSHR). We constructed a mutant of eelFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 614 (eelFSHR-t614). The eelFSHR-t614 lacked all potential phosphorylation sites present in the C-terminal region of eelFSHR. In order to obtain the eelFSHR ligand, we produced recombinant follicle-stimulating hormone (rec-eelFSHβ/α) in the CHO-suspension cells. The expression level was 2-3 times higher than that of the transient expression of eelFSH in attached CHO-K1 cells. The molecular weight of the rec-eelFSHβ/α protein was identified to be approximately 34 kDa. The cells expressing eelFSHR-t614 showed an increase in agonist-induced cAMP responsiveness. The maximal cAMP responses of cells expressing eelFSHR-t614 were lower than those of c...Continue Reading

References

Jan 28, 1999·Molecular and Cellular Endocrinology·J S TapanainenI T Huhtaniemi
Aug 9, 2003·Molecular Endocrinology·Hanumanthappa KrishnamurthyMario Ascoli
Jul 1, 2004·The Journal of Reproduction and Development·Kwan-Sik MinKunio Shiota
Mar 29, 2008·Journal of Molecular Endocrinology·Sébastien LegardinierClaire Cahoreau
Aug 10, 2017·Balsaeng'gwa saengsig·Jong-Ju ParkKwan-Sik Min
Mar 11, 2018·The Journal of Biological Chemistry·Qingwen WanNevin A Lambert
May 13, 2018·The Journal of Biological Chemistry·Kirill A Martemyanov, Mikel Garcia-Marcos

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Methods Mentioned

BETA
PCR
transfection
enzyme-linked immunosorbent assay
ELISA
electrophoresis
X-ray

Software Mentioned

PathHunter
GraFit
GraphPad
GraphPad Prism

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