PMID- 10585874
OWN - NLM
STAT- MEDLINE
DCOM- 20000317
LR  - 20181113
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 344 Pt 3
DP  - 1999 Dec 15
TI  - Direct interaction between p47phox and protein kinase C: evidence for targeting
      of protein kinase C by p47phox in neutrophils.
PG  - 859-66
AB  - p47(phox) is an essential component of the NADPH oxidase, and phosphorylation of 
      p47(phox) is associated with activation of the enzyme. Here we have used
      p47(phox) affinity chromatography to extract a p47(phox) kinase from neutrophil
      cytosol. The kinase activity was purified by gel filtration and Mini Q
      chromatography and shown to be indistinguishable from the catalytic fragments of 
      protein kinase C (PKC)-beta(I), -beta(II) and -delta. The C-terminus of p47(phox)
      represented the site of interaction with PKC. Co-immunoprecipitation experiments 
      revealed that the interaction between PKC isotypes and p47(phox) takes place in
      intact cells. However PKC-beta and -delta showed different time courses of
      co-immunoprecipitation, suggesting that the interactions may serve different
      functions for the various PKC isotypes. Using cells lacking p47(phox), we
      investigated the functional relevance of the interaction between PKC and
      p47(phox). Subcellular fractionation revealed an abnormal recruitment of
      PKC-beta(I) and -beta(II), but not PKC-delta, to particulate fractions in
      p47(phox)-deficient cells. Phosphorylation of cytosolic proteins was generally
      increased in stimulated p47(phox)-deficient neutrophils as compared with normal
      neutrophils. Furthermore, the cytoskeletal protein coronin was not phosphorylated
      upon stimulation of p47(phox)-deficient neutrophils. These findings were
      confirmed in an in vitro-reconstituted system using rat brain cytosol in which
      addition of p47(phox) affected phosphorylation by PKC/PKM (PKM is the catalytic
      fragment of PKC). These results indicate that p47(phox) can act as a regulator of
      PKC in neutrophils.
FAU - Reeves, E P
AU  - Reeves EP
AD  - Centre for Molecular Medicine, University College London, 5 University Street,
      London WC1E 6JJ, U.K.
FAU - Dekker, L V
AU  - Dekker LV
FAU - Forbes, L V
AU  - Forbes LV
FAU - Wientjes, F B
AU  - Wientjes FB
FAU - Grogan, A
AU  - Grogan A
FAU - Pappin, D J
AU  - Pappin DJ
FAU - Segal, A W
AU  - Segal AW
LA  - eng
GR  - Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Microfilament Proteins)
RN  - 0 (Phosphopeptides)
RN  - 0 (Phosphoproteins)
RN  - 0 (Protein Isoforms)
RN  - 145420-64-0 (coronin proteins)
RN  - EC 1.6.3.- (NADPH Oxidases)
RN  - EC 1.6.3.1 (neutrophil cytosolic factor 1)
RN  - EC 2.7.11.13 (Protein Kinase C)
SB  - IM
EIN - Biochem J 2000 Feb 1;345 Pt 3:767
MH  - Animals
MH  - Brain/metabolism
MH  - Cell Fractionation
MH  - Cytosol/metabolism
MH  - Humans
MH  - Microfilament Proteins/metabolism
MH  - NADPH Oxidases
MH  - Neutrophils/enzymology/*metabolism
MH  - Phosphopeptides/chemistry
MH  - Phosphoproteins/*metabolism
MH  - Phosphorylation
MH  - Protein Binding
MH  - Protein Isoforms
MH  - Protein Kinase C/*metabolism
MH  - Rats
PMC - PMC1220709
EDAT- 1999/12/10 09:00
MHDA- 2000/05/29 09:00
CRDT- 1999/12/10 09:00
PHST- 1999/12/10 09:00 [pubmed]
PHST- 2000/05/29 09:00 [medline]
PHST- 1999/12/10 09:00 [entrez]
PST - ppublish
SO  - Biochem J. 1999 Dec 15;344 Pt 3:859-66.