|
KMID : 1161320200350020155
|
|
Journal of Animal Reproduciton and Biotechnology
2020 Volume.35 No. 2 p.155 ~ p.162
|
|
|
Signal transduction of C-terminal phosphorylation sites for equine follicle stimulating hormone receptor (eFSHR)
|
|
Seong Hoon-Ki
Choi Seung-Hee
Byambaragchaa Munkhzaya
Min Kwan-Sik
|
|
|
Abstract
|
|
|
|
Equine follicle stimulating hormone receptor (eFSHR) has a large extracellular domain and an intracellular domain containing approximately 10 phosphorylation sites within the G protein-coupled receptor. This study was conducted to analyze the function of phosphorylation sties at the eFSHR C-terminal region. We constructed a mutant of eFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 641 (eFSHR-t641). This removed 10 potential phosphorylation sites from the C-terminal region of the intracellular loop. The eFSHR-wild type (eFSHR-wt) and eFSHR-t641 cDNAs were subcloned into the pCMV-ARMS1-PK2 expression vector. These plasmids were transfected into PathHunter CHO-K1 Parental cells expressing β-arrestin 2 enzyme acceptor fusion protein and analyzed for agonist-induced cAMP response. The cAMP response in cells expressing eFSHR-t641 was lower than the response in cells expressing eFSHR-wt. EC50 values of eFSHR-wt and eFSHR-t641 were 1079 ng/mL and 1834 ng/mL, respectively. eFSHR-t641 was approximately 0.58-fold compared with that of eFSHR-wt. The maximal response in eFSHR-wt and eFSHR-t641 was 24.7 nM and 16.7 nM, respectively. The Rmax value of phosphorylation sites in eFSHR-t641 was also decreased to approximately 68.4% of that in eFSHR-wt. The collective data implicate that the phosphorylation sites in the eFSHR C-terminal region have a pivotal role in signal transduction in PathHunter CHO-K1 cells, and indicate that β-arrestin is involved in coupling the activated receptors to the internalization system.
|
|
|
KEYWORD
|
|
|
eFSHR, PathHunter CHO-K1 Parental cells, phosphorylation sites, signal transduction
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|