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PMID: 8770870 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The cytosolic subunit p67phox contains an NADPH-binding site that participates in catalysis by the leukocyte NADPH oxidase.

The Journal of clinical investigation ·Vol. 98 ·No. 4 ·1996-08-15 ·Pages 977-83

Smith RM, Connor JA, Chen LM, Babior BM

Abstract

The NADPH-dependent respiratory burst oxidase of human neutrophils catalyzes the reduction of oxygen to superoxide using NADPH as the electron donor and is essential for normal host defenses. To gain insight into the function of the various oxidase subunits that are required for the full expression of catalytic activity, we studied the interactions between the 2',3'-dialdehyde derivative of NADPH (NADPH dialdehyde) and neutrophil cytosol. NADPH dialdehyde treatment of cytosol resulted in the loss of the ability of the cytosol to participate in cell-free oxidase activation; this inactivation was blocked by NADPH but not by NAD, NADP, or GTP. Partial purification of neutrophil cytosol yielded a single peak which could restore the activity lost in cytosol treated with NADPH dialdehyde. This peak contained p67phox but not p47phox or Rac2. Purified recombinant p67phox was similarly able to restore the activity lost in NADPH dialdehyde-treated cytosol and bound [32P]NADPH dialdehyde in a specific fashion. The activity of recombinant p67phox in cell-free oxidase assays was lost on treatment with NADPH dialdehyde. Together, these data suggest p67phox contains the catalytic NADPH-binding site of the leukocyte NADPH oxidase.

MeSH Terms
Aldehydes/chemistry Amino Acid Sequence Binding Sites Catalysis Cytosol/enzymology Humans Molecular Sequence Data NADH, NADPH Oxidoreductases/chemistry,metabolism NADP/chemistry,metabolism NADPH Oxidases Neutrophils/enzymology Phosphoproteins/metabolism Respiratory Burst
Chemicals
Aldehydes Phosphoproteins neutrophil cytosol factor 67K NADP NADH, NADPH Oxidoreductases NADPH Oxidases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Smith R M
The Department of Medicine, University of California, San Diego 92093, USA.
Connor J A
Chen L M
Babior B M
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Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI118882
Published
1996-08-15
Pages
977-83
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC507513
Subset
IM
Grants
NIAID NIH HHS · AI24227 · United States
NIAID NIH HHS · AI30742 · United States
NHLBI NIH HHS · HL23584 · United States
External Links
PubMed source DOI: 10.1172/JCI118882 PMC Full Text View journal details
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