PMID- 10567430
OWN - NLM
STAT- MEDLINE
DCOM- 19991229
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 48
DP  - 1999 Nov 26
TI  - Selective regulation of Galpha(q/11) by an RGS domain in the G protein-coupled
      receptor kinase, GRK2.
PG  - 34483-92
AB  - G protein-coupled receptor kinases (GRKs) are well characterized regulators of G 
      protein-coupled receptors, whereas regulators of G protein signaling (RGS)
      proteins directly control the activity of G protein alpha subunits.
      Interestingly, a recent report (Siderovski, D. P., Hessel, A., Chung, S., Mak, T.
      W., and Tyers, M. (1996) Curr. Biol. 6, 211-212) identified a region within the N
      terminus of GRKs that contained homology to RGS domains. Given that RGS domains
      demonstrate AlF(4)(-)-dependent binding to G protein alpha subunits, we tested
      the ability of G proteins from a crude bovine brain extract to bind to GRK
      affinity columns in the absence or presence of AlF(4)(-). This revealed the
      specific ability of bovine brain Galpha(q/11) to bind to both GRK2 and GRK3 in an
      AlF(4)(-)-dependent manner. In contrast, Galpha(s), Galpha(i), and Galpha(12/13) 
      did not bind to GRK2 or GRK3 despite their presence in the extract. Additional
      studies revealed that bovine brain Galpha(q/11) could also bind to an N-terminal 
      construct of GRK2, while no binding of Galpha(q/11), Galpha(s), Galpha(i), or
      Galpha(12/13) to comparable constructs of GRK5 or GRK6 was observed. Experiments 
      using purified Galpha(q) revealed significant binding of both Galpha(q)
      GDP/AlF(4)(-) and Galpha(q)(GTPgammaS), but not Galpha(q)(GDP), to GRK2.
      Activation-dependent binding was also observed in both COS-1 and HEK293 cells as 
      GRK2 significantly co-immunoprecipitated constitutively active Galpha(q)(R183C)
      but not wild type Galpha(q). In vitro analysis revealed that GRK2 possesses weak 
      GAP activity toward Galpha(q) that is dependent on the presence of a G
      protein-coupled receptor. However, GRK2 effectively inhibited Galpha(q)-mediated 
      activation of phospholipase C-beta both in vitro and in cells, possibly through
      sequestration of activated Galpha(q). These data suggest that a subfamily of the 
      GRKs may be bifunctional regulators of G protein-coupled receptor signaling
      operating directly on both receptors and G proteins.
FAU - Carman, C V
AU  - Carman CV
AD  - Department of Biochemistry, Kimmel Cancer Institute, Thomas Jefferson University,
      Philadelphia, Pennsylvania 19107, USA.
FAU - Parent, J L
AU  - Parent JL
FAU - Day, P W
AU  - Day PW
FAU - Pronin, A N
AU  - Pronin AN
FAU - Sternweis, P M
AU  - Sternweis PM
FAU - Wedegaertner, P B
AU  - Wedegaertner PB
FAU - Gilman, A G
AU  - Gilman AG
FAU - Benovic, J L
AU  - Benovic JL
FAU - Kozasa, T
AU  - Kozasa T
LA  - eng
GR  - 5-T32-CA09662/CA/NCI NIH HHS/United States
GR  - GM34497/GM/NIGMS NIH HHS/United States
GR  - GM44944/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Aluminum Compounds)
RN  - 0 (Isoenzymes)
RN  - 0 (RGS Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Tissue Extracts)
RN  - 21340-02-3 (tetrafluoroaluminate)
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 (ADRBK2 protein, human)
RN  - EC 2.7.11.15 (G-Protein-Coupled Receptor Kinase 3)
RN  - EC 2.7.11.15 (beta-Adrenergic Receptor Kinases)
RN  - EC 3.1.4.- (Type C Phospholipases)
RN  - EC 3.1.4.11 (Phospholipase C beta)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
RN  - EC 3.6.5.1 (GTP-Binding Protein alpha Subunits, Gq-G11)
RN  - Q80VPU408O (Fluorides)
SB  - IM
MH  - Aluminum Compounds/pharmacology
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites/physiology
MH  - Binding, Competitive
MH  - Brain/metabolism
MH  - COS Cells
MH  - Cattle
MH  - Cell Line
MH  - Cyclic AMP-Dependent Protein Kinases/chemistry/genetics/*metabolism
MH  - Enzyme Activation
MH  - Fluorides/pharmacology
MH  - G-Protein-Coupled Receptor Kinase 3
MH  - GTP-Binding Protein alpha Subunits, Gq-G11
MH  - GTP-Binding Proteins/genetics/*metabolism
MH  - Humans
MH  - Isoenzymes/metabolism
MH  - Kinetics
MH  - Molecular Sequence Data
MH  - Phospholipase C beta
MH  - Protein Binding/drug effects
MH  - Protein Structure, Tertiary
MH  - Protein-Serine-Threonine Kinases/genetics/metabolism
MH  - RGS Proteins/chemistry/*metabolism
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Tissue Extracts/metabolism
MH  - Type C Phospholipases/metabolism
MH  - beta-Adrenergic Receptor Kinases
EDAT- 1999/11/24 00:00
MHDA- 1999/11/24 00:01
CRDT- 1999/11/24 00:00
PHST- 1999/11/24 00:00 [pubmed]
PHST- 1999/11/24 00:01 [medline]
PHST- 1999/11/24 00:00 [entrez]
AID - 10.1074/jbc.274.48.34483 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Nov 26;274(48):34483-92. doi: 10.1074/jbc.274.48.34483.