PMID- 10446210
OWN - NLM
STAT- MEDLINE
DCOM- 19990909
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 34
DP  - 1999 Aug 20
TI  - G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger
      regulatory factor via a PDZ domain-mediated interaction.
PG  - 24328-34
AB  - The Na(+)/H(+) exchanger regulatory factor (NHERF) is constitutively
      phosphorylated in cells, but the site(s) of this phosphorylation and the
      kinase(s) responsible for it have not been identified. We show here that the
      primary site of constitutive NHERF phosphorylation in human embryonic kidney 293 
      (HEK-293) cells is Ser(289), and that the stoichiometry of phosphorylation is
      near 1 mol/mol. NHERF contains two PDZ domains that recognize the sequence
      S/T-X-L at the carboxyl terminus of target proteins, and thus we examined the
      possibility that kinases containing this motif might associate with and
      phosphorylate NHERF. Overlay experiments and co-immunoprecipitation studies
      revealed that NHERF binds with high affinity to a splice variant of the G
      protein-coupled receptor kinase 6, GRK6A, which terminates in the motif T-R-L.
      NHERF does not associate with GRK6B or GRK6C, alternatively spliced variants that
      differ from GRK6A at their extreme carboxyl termini. GRK6A phosphorylates NHERF
      efficiently on Ser(289) in vitro, whereas GRK6B, GRK6C, and GRK2 do not.
      Furthermore, the endogenous "NHERF kinase" activity in HEK-293 cell lysates is
      sensitive to treatments that alter the activity of GRK6A. These data suggest that
      GRK6A phosphorylates NHERF via a PDZ domain-mediated interaction and that GRK6A
      is the kinase in HEK-293 cells responsible for the constitutive phosphorylation
      of NHERF.
FAU - Hall, R A
AU  - Hall RA
AD  - Howard Hughes Medical Institute, Departments of Medicine and Biochemistry, Duke
      University Medical Center, Durham, North Carolina 27710, USA.
FAU - Spurney, R F
AU  - Spurney RF
FAU - Premont, R T
AU  - Premont RT
FAU - Rahman, N
AU  - Rahman N
FAU - Blitzer, J T
AU  - Blitzer JT
FAU - Pitcher, J A
AU  - Pitcher JA
FAU - Lefkowitz, R J
AU  - Lefkowitz RJ
LA  - eng
GR  - HL16037/HL/NHLBI 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 (Phosphoproteins)
RN  - 0 (Sodium-Hydrogen Exchangers)
RN  - 0 (sodium-hydrogen exchanger regulatory factor)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.16 (G-Protein-Coupled Receptor Kinases)
RN  - EC 2.7.11.16 (G-protein-coupled receptor kinase 6)
RN  - H88EPA0A3N (Staurosporine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Cells, Cultured
MH  - G-Protein-Coupled Receptor Kinases
MH  - Humans
MH  - Molecular Sequence Data
MH  - Phosphoproteins/*metabolism
MH  - Phosphorylation
MH  - *Protein-Serine-Threonine Kinases
MH  - Receptor Protein-Tyrosine Kinases/chemistry/*physiology
MH  - Sodium-Hydrogen Exchangers/metabolism
MH  - Staurosporine/pharmacology
EDAT- 1999/08/14 00:00
MHDA- 1999/08/14 00:01
CRDT- 1999/08/14 00:00
PHST- 1999/08/14 00:00 [pubmed]
PHST- 1999/08/14 00:01 [medline]
PHST- 1999/08/14 00:00 [entrez]
AID - 10.1074/jbc.274.34.24328 [doi]
AID - S0021-9258(19)55563-6 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Aug 20;274(34):24328-34. doi: 10.1074/jbc.274.34.24328.