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PMID: 11248021 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Assembly of the neutrophil respiratory burst oxidase: a direct interaction between p67PHOX and cytochrome b558.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 98 ·No. 6 ·2001-03-13 ·Pages 3001-5

Dang PM, Cross AR, Babior BM

Abstract

Activation of the phagocyte NADPH oxidase complex requires the assembly of the cytosolic factors p47(PHOX), p67(PHOX), p40(PHOX), and Rac1 or Rac2, with the membrane-bound cytochrome b(558). Whereas the interaction of p47(PHOX) with cytochrome b(558) is well established, an interaction between p67(PHOX) and cytochrome b(558) has never been investigated. We report here a direct interaction between p67(PHOX) and cytochrome b(558). First, labeled p67(PHOX) recognizes a 91-kDa band in specific granules from a normal patient but not from a cytochrome b(558)-deficient patient. Second, p67(PHOX) binds to cytochrome b(558) that has been bound to nitrocellulose. Third, GTP-p67(PHOX) bound to glutathione agarose is able to pull down cytochrome b(558.) Rac1-GTP or Rac1-GDP increased the binding of p67(PHOX) to cytochrome b(558), suggesting that at least one of the oxidase-related functions of Rac1 is to promote the interaction between p67(PHOX) and cytochrome b(558).

MeSH Terms
Cytochrome b Group/metabolism Guanosine 5'-O-(3-Thiotriphosphate)/metabolism Humans Membrane Glycoproteins/metabolism NADH, NADPH Oxidoreductases/metabolism NADPH Oxidase 2 NADPH Oxidases/genetics,metabolism Neutrophils/enzymology Phosphoproteins/genetics,metabolism Recombinant Fusion Proteins/genetics,metabolism Respiratory Burst rac1 GTP-Binding Protein/metabolism
Chemicals
Cytochrome b Group Membrane Glycoproteins Phosphoproteins Recombinant Fusion Proteins neutrophil cytosol factor 67K Guanosine 5'-O-(3-Thiotriphosphate) cytochrome b558 NADH, NADPH Oxidoreductases CYBB protein, human NADPH Oxidase 2 NADPH Oxidases superoxide-forming enzyme rac1 GTP-Binding Protein
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Dang P M
Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA.
Cross A R
Babior B M
References (22)
22 references, click to expand
  1. Binding of nicotinamide adenine dinucleotide phosphate to the tetratricopeptide repeat domains at the N-terminus of p67PHOX, a subunit of the leukocyte nicotinamide adenine dinucleotide phosphate oxidase.
    Biochemistry. 2000 Mar 21;39(11):3069-75 PMID: 10715128
  2. The p67(phox) activation domain regulates electron flow from NADPH to flavin in flavocytochrome b(558).
    J Biol Chem. 1999 Aug 13;274(33):22999-3005 PMID: 10438466
  3. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  4. A possible role for protein phosphorylation in the activation of the respiratory burst in human neutrophils. Evidence from studies with cells from patients with chronic granulomatous disease.
    J Biol Chem. 1986 Jul 15;261(20):9109-15 PMID: 3087987
  5. Cytochrome b-245 is a flavocytochrome containing FAD and the NADPH-binding site of the microbicidal oxidase of phagocytes.
    Biochem J. 1992 Jun 15;284 ( Pt 3):781-8 PMID: 1320378
  6. Requirement for posttranslational processing of Rac GTP-binding proteins for activation of human neutrophil NADPH oxidase.
    Mol Biol Cell. 1993 Mar;4(3):261-9 PMID: 8387355
  7. Cytosolic guanine nucleotide-binding protein Rac2 operates in vivo as a component of the neutrophil respiratory burst oxidase. Transfer of Rac2 and the cytosolic oxidase components p47phox and p67phox to the submembranous actin cytoskeleton during oxidase activation.
    J Biol Chem. 1994 Mar 4;269(9):6729-34 PMID: 8120032
  8. Isolation and characterization of gelatinase granules from human neutrophils.
    Blood. 1994 Mar 15;83(6):1640-9 PMID: 8123855
  9. Role of Src homology 3 domains in assembly and activation of the phagocyte NADPH oxidase.
    Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5345-9 PMID: 8202490
  10. Peptides corresponding to the region adjacent to His-94 in the small subunit of cytochrome b558 inhibit superoxide generation in a cell-free system from human neutrophils.
    Biochem Biophys Res Commun. 1994 Oct 28;204(2):924-9 PMID: 7980562
  11. Function of wild-type or mutant Rac2 and Rap1a GTPases in differentiated HL60 cell NADPH oxidase activation.
    Blood. 1995 Feb 1;85(3):804-11 PMID: 7833480
  12. Functional reconstitution of the phagocyte NADPH oxidase by transfection of its multiple components in a heterologous system.
    Blood. 1995 Feb 15;85(4):1104-10 PMID: 7849298
  13. Mapping the domains of interaction of p40phox with both p47phox and p67phox of the neutrophil oxidase complex using the two-hybrid system.
    J Biol Chem. 1995 Mar 17;270(11):5695-7 PMID: 7890694
  14. The cytosolic subunit p67phox contains an NADPH-binding site that participates in catalysis by the leukocyte NADPH oxidase.
    J Clin Invest. 1996 Aug 15;98(4):977-83 PMID: 8770870
  15. The cytosolic component p47(phox) is not a sine qua non participant in the activation of NADPH oxidase but is required for optimal superoxide production.
    J Biol Chem. 1996 Nov 29;271(48):30326-9 PMID: 8939991
  16. Involvement of p40phox in activation of phagocyte NADPH oxidase through association of its carboxyl-terminal, but not its amino-terminal, with p67phox.
    J Exp Med. 1996 Sep 1;184(3):893-902 PMID: 9064349
  17. Synthetic peptides corresponding to various hydrophilic regions of the large subunit of cytochrome b558 inhibit superoxide generation in a cell-free system from neutrophils.
    Biochem Biophys Res Commun. 1997 May 19;234(2):531-6 PMID: 9177307
  18. Regulation of the neutrophil respiratory burst oxidase. Identification of an activation domain in p67(phox).
    J Biol Chem. 1998 Jul 3;273(27):16663-8 PMID: 9642219
  19. Spontaneous activation of NADPH oxidase in a cell-free system: unexpected multiple effects of magnesium ion concentrations.
    Biochem J. 1999 Feb 15;338 ( Pt 1):229-33 PMID: 9931320
  20. NADPH oxidase: an update.
    Blood. 1999 Mar 1;93(5):1464-76 PMID: 10029572
  21. Activation of the phagocyte NADPH oxidase protein p47(phox). Phosphorylation controls SH3 domain-dependent binding to p22(phox).
    J Biol Chem. 1999 Jul 9;274(28):19731-7 PMID: 10391914
  22. p40(phox) Participates in the activation of NADPH oxidase by increasing the affinity of p47(phox) for flavocytochrome b(558).
    Biochem J. 2000 Jul 1;349(Pt 1):113-7 PMID: 10861218
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.061029698
Published
2001-03-13
Pages
3001-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC30596
Subset
IM
Grants
NIAID NIH HHS · R01 AI024838 · United States
NIAID NIH HHS · R01 AI024227 · United States
NIAID NIH HHS · AI-24838 · United States
NIAID NIH HHS · AI-28479 · United States
NIAID NIH HHS · AI-24227 · United States
NIAID NIH HHS · R01 AI028479 · United States
External Links
PubMed source DOI: 10.1073/pnas.061029698 PMC Full Text View journal details
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