PMID- 10334944
OWN - NLM
STAT- MEDLINE
DCOM- 19990625
LR  - 20131121
IS  - 0006-291X (Print)
IS  - 0006-291X (Linking)
VI  - 259
IP  - 1
DP  - 1999 May 27
TI  - Overexpression of wild-type and catalytically inactive forms of GRK2 and GRK6
      fails to alter the agonist-induced phosphorylation of the C5a receptor (CD88):
      evidence that GRK6 is autophosphorylated in COS-7 cells.
PG  - 224-9
AB  - The G protein-coupled receptor kinase family comprises six members (GRK1 to GRK6)
      that phosphorylate and desensitize a number of agonist-occupied G protein-coupled
      receptors. Overexpression of the dominant negative mutant GRK2-K220R is often
      accompanied by an inhibition of the agonist-mediated phosphorylation of G
      protein-coupled receptors. In the case of the C5a receptor (C5aR), the
      overexpression of wild-type GRK2 or GRK6 as well as of catalytically inactive
      forms of these kinases (GRK2-K220R and GRK6-K215R) failed to increase or to
      inhibit the agonist-mediated phosphorylation of C5aR, respectively. Replacement
      of Lys215 by an arginine residue in GRK6 yielded a protein with a relative
      molecular mass of 63 kDa, whereas wild-type GRK6 had a relative molecular mass of
      66 kDa on polyacrylamide gel. The mutations S484D and T485D in the catalytically 
      inactive mutant GRK6-K215R resulted in a protein (GRK6-RDD) with the same
      electrophoretic mobility as wild-type GRK6. Furthermore, in the absence of
      phosphatase inhibitors, GRK6 was rapidly converted into the 63 kDa species,
      whereas GRK6-RDD was not. Overepression of GRK6-RDD failed to alter the
      agonist-mediated phosphorylation of C5aR. Taken together, the results suggest
      that C5aR is not a substrate for either GRK2 or GRK6 and that GRK6 is very likely
      autophosphorylated on Ser484 and Thr485 in vivo.
CI  - Copyright 1999 Academic Press.
FAU - Milcent, M D
AU  - Milcent MD
AD  - DBMS/Laboratoire de Biochimie et de Biophysique des Systemes Integres (UMR 314
      CEA/CNRS), CEA/Grenoble, 17 rue des Martyrs, Grenoble Cedex 9, 38054, France.
FAU - Christophe, T
AU  - Christophe T
FAU - Rabiet, M J
AU  - Rabiet MJ
FAU - Tardif, M
AU  - Tardif M
FAU - Boulay, F
AU  - Boulay F
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 0 (Antigens, CD)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Receptor, Anaphylatoxin C5a)
RN  - 0 (Receptors, Complement)
RN  - 0 (Recombinant Proteins)
RN  - 1W21G5Q4N2 (Okadaic Acid)
RN  - 80295-54-1 (Complement C5a)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases)
RN  - EC 2.7.11.15 (beta-Adrenergic Receptor Kinases)
RN  - EC 2.7.11.16 (G-Protein-Coupled Receptor Kinases)
RN  - EC 2.7.11.16 (G-protein-coupled receptor kinase 6)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antigens, CD/*metabolism
MH  - COS Cells
MH  - Complement C5a/genetics/metabolism
MH  - Cyclic AMP-Dependent Protein Kinases/*genetics
MH  - Enzyme Inhibitors/pharmacology
MH  - G-Protein-Coupled Receptor Kinases
MH  - Gene Expression/genetics
MH  - Humans
MH  - Kinetics
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Mutation/genetics
MH  - Okadaic Acid/pharmacology
MH  - Phosphorylation
MH  - *Protein-Serine-Threonine Kinases
MH  - Receptor Protein-Tyrosine Kinases/*genetics
MH  - Receptor, Anaphylatoxin C5a
MH  - Receptors, Complement/*metabolism
MH  - Recombinant Proteins/metabolism
MH  - Transfection
MH  - beta-Adrenergic Receptor Kinases
EDAT- 1999/05/21 00:00
MHDA- 1999/05/21 00:01
CRDT- 1999/05/21 00:00
PHST- 1999/05/21 00:00 [pubmed]
PHST- 1999/05/21 00:01 [medline]
PHST- 1999/05/21 00:00 [entrez]
AID - 10.1006/bbrc.1999.0758 [doi]
AID - S0006-291X(99)90758-1 [pii]
PST - ppublish
SO  - Biochem Biophys Res Commun. 1999 May 27;259(1):224-9. doi:
      10.1006/bbrc.1999.0758.