PMID- 23106126
OWN - NLM
STAT- MEDLINE
DCOM- 20130306
LR  - 20220129
IS  - 1471-4159 (Electronic)
IS  - 0022-3042 (Print)
IS  - 0022-3042 (Linking)
VI  - 124
IP  - 2
DP  - 2013 Jan
TI  - Identification of phosphorylation sites in the COOH-terminal tail of the mu-opioid 
      receptor.
PG  - 189-99
LID - 10.1111/jnc.12071 [doi]
AB  - Phosphorylation is considered a key event in the signalling and regulation of the 
      mu opioid receptor (MOPr). Here, we used mass spectroscopy to determine the 
      phosphorylation status of the C-terminal tail of the rat MOPr expressed in human 
      embryonic kidney 293 (HEK-293) cells. Under basal conditions, MOPr is 
      phosphorylated on Ser(363) and Thr(370), while in the presence of morphine or 
      [D-Ala2, NMe-Phe4, Gly-ol5]-enkephalin (DAMGO), the COOH terminus is 
      phosphorylated at three additional residues, Ser(356) , Thr(357) and Ser(375). 
      Using N-terminal glutathione S transferase (GST) fusion proteins of the 
      cytoplasmic, C-terminal tail of MOPr and point mutations of the same, we show 
      that, in vitro, purified G protein-coupled receptor kinase 2 (GRK2) 
      phosphorylates Ser(375), protein kinase C (PKC) phosphorylates Ser(363), while 
      CaMKII phosphorylates Thr(370). Phosphorylation of the GST fusion protein of the 
      C-terminal tail of MOPr enhanced its ability to bind arrestin-2 and -3. Hence, 
      our study identifies both the basal and agonist-stimulated phospho-acceptor sites 
      in the C-terminal tail of MOPr, and suggests that the receptor is subject to 
      phosphorylation and hence regulation by multiple protein kinases.
CI  - (c) 2012 International Society for Neurochemistry.
FAU - Chen, Ying-Ju
AU  - Chen YJ
AD  - School of Physiology and Pharmacology, University of Bristol, Bristol, UK.
FAU - Oldfield, Sue
AU  - Oldfield S
FAU - Butcher, Adrian J
AU  - Butcher AJ
FAU - Tobin, Andrew B
AU  - Tobin AB
FAU - Saxena, Kunal
AU  - Saxena K
FAU - Gurevich, Vsevolod V
AU  - Gurevich VV
FAU - Benovic, Jeffrey L
AU  - Benovic JL
FAU - Henderson, Graeme
AU  - Henderson G
FAU - Kelly, Eamonn
AU  - Kelly E
LA  - eng
GR  - G1100712/MRC_/Medical Research Council/United Kingdom
GR  - MR/J013269/1/MRC_/Medical Research Council/United Kingdom
GR  - R01 GM077561/GM/NIGMS NIH HHS/United States
GR  - GM077561/GM/NIGMS NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20121130
PL  - England
TA  - J Neurochem
JT  - Journal of neurochemistry
JID - 2985190R
RN  - 0 (Receptors, Opioid, mu)
RN  - EC 2.7.- (Protein Kinases)
SB  - IM
MH  - 5' Flanking Region/genetics
MH  - Amino Acid Sequence
MH  - Animals
MH  - HEK293 Cells
MH  - Humans
MH  - Molecular Sequence Data
MH  - Phosphorylation/genetics
MH  - Protein Kinases/genetics/metabolism
MH  - Protein Structure, Tertiary/genetics
MH  - Protein Transport/genetics
MH  - Rats
MH  - Receptors, Opioid, mu/*chemistry/*genetics/metabolism
PMC - PMC4226418
MID - NIHMS417657
EDAT- 2012/10/31 06:00
MHDA- 2013/03/07 06:00
PMCR- 2014/11/10
CRDT- 2012/10/31 06:00
PHST- 2012/08/10 00:00 [received]
PHST- 2012/10/10 00:00 [revised]
PHST- 2012/10/18 00:00 [accepted]
PHST- 2012/10/31 06:00 [entrez]
PHST- 2012/10/31 06:00 [pubmed]
PHST- 2013/03/07 06:00 [medline]
PHST- 2014/11/10 00:00 [pmc-release]
AID - 10.1111/jnc.12071 [doi]
PST - ppublish
SO  - J Neurochem. 2013 Jan;124(2):189-99. doi: 10.1111/jnc.12071. Epub 2012 Nov 30.