PMID- 9202043
OWN - NLM
STAT- MEDLINE
DCOM- 19970731
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 272
IP  - 27
DP  - 1997 Jul 4
TI  - Phosphorylation of the respiratory burst oxidase subunit p67(phox) during human 
      neutrophil activation. Regulation by protein kinase C-dependent and independent 
      pathways.
PG  - 17204-8
AB  - The respiratory burst oxidase of phagocytes and B lymphocytes catalyzes the 
      reduction of oxygen to superoxide anion (O-2) at the expense of NADPH. This 
      multicomponent enzyme is dormant in resting cells but is activated on exposure to 
      an appropriate stimulus. The phosphorylation-dependent mechanisms regulating the 
      activation of the respiratory burst oxidase are unclear, particularly the 
      phosphorylation status of the cytosolic component p67(phox). In this study, we 
      found that activation of human neutrophils with 
      formyl-methionyl-leucyl-phenylalanine (fMLP), a chemotactic peptide, or phorbol 
      myristate acetate (PMA), a stimulator of protein kinase C (PKC), resulted in the 
      phosphorylation of p67(phox). Using an anti-p67(phox) antibody or an 
      anti-p47(phox) antibody, we showed that phosphorylated p67(phox) and p47(phox) 
      form a complex. Phosphoamino acid analysis of the phosphorylated p67(phox) 
      revealed only 32P-labeled serine residues. Two-dimensional tryptic peptide 
      mapping analysis showed that p67(phox) is phosphorylated at the same peptide 
      whether fMLP or PMA is used as a stimulus. In addition, PKC induced the 
      phosphorylation of recombinant GST-p67(phox) in vitro, at the same peptide as 
      that phosphorylated in intact cells. PMA-induced phosphorylation of p67(phox) was 
      strongly inhibited by the PKC inhibitor GF109203X. In contrast, fMLP-induced 
      phosphorylation was minimally affected by this PKC inhibitor. Taken together, 
      these results show that p67(phox) is phosphorylated in human neutrophils by 
      different pathways, one of which involves protein kinase C.
FAU - Benna, J E
AU  - Benna JE
AD  - INSERM U294, CHU Xavier Bichat, Service d'Hematologie et d'Immunologie 
      Biologiques, 46 rue Henri Huchard, 75018 Paris, France. benna@bichat.inserm.fr
FAU - Dang, P M
AU  - Dang PM
FAU - Gaudry, M
AU  - Gaudry M
FAU - Fay, M
AU  - Fay M
FAU - Morel, F
AU  - Morel F
FAU - Hakim, J
AU  - Hakim J
FAU - Gougerot-Pocidalo, M A
AU  - Gougerot-Pocidalo MA
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Indoles)
RN  - 0 (Maleimides)
RN  - 0 (Phosphoproteins)
RN  - 0 (neutrophil cytosol factor 67K)
RN  - 452VLY9402 (Serine)
RN  - 59880-97-6 (N-Formylmethionine Leucyl-Phenylalanine)
RN  - EC 1.6.- (NADH, NADPH Oxidoreductases)
RN  - EC 1.6.3.- (NADPH Oxidases)
RN  - EC 1.6.3.1 (neutrophil cytosolic factor 1)
RN  - EC 1.6.99.- (superoxide-forming enzyme)
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - L79H6N0V6C (bisindolylmaleimide I)
RN  - NI40JAQ945 (Tetradecanoylphorbol Acetate)
SB  - IM
MH  - Enzyme Activation
MH  - Enzyme Inhibitors/pharmacology
MH  - Humans
MH  - Indoles/pharmacology
MH  - *Lymphocyte Activation
MH  - Maleimides/pharmacology
MH  - N-Formylmethionine Leucyl-Phenylalanine/pharmacology
MH  - NADH, NADPH Oxidoreductases/*metabolism
MH  - NADPH Oxidases/metabolism
MH  - Neutrophils/*metabolism
MH  - Peptide Mapping
MH  - Phosphoproteins/*metabolism
MH  - Phosphorylation
MH  - Protein Kinase C/antagonists & inhibitors/*metabolism
MH  - Serine/metabolism
MH  - Tetradecanoylphorbol Acetate/pharmacology
EDAT- 1997/07/04 00:00
MHDA- 1997/07/04 00:01
CRDT- 1997/07/04 00:00
PHST- 1997/07/04 00:00 [pubmed]
PHST- 1997/07/04 00:01 [medline]
PHST- 1997/07/04 00:00 [entrez]
AID - S0021-9258(18)39352-9 [pii]
AID - 10.1074/jbc.272.27.17204 [doi]
PST - ppublish
SO  - J Biol Chem. 1997 Jul 4;272(27):17204-8. doi: 10.1074/jbc.272.27.17204.