Two cytosolic components of the neutrophil NADPH oxidase, P47-PHOX and P67-PHOX, are not flavoproteins

Tsukasa Chiba, Mizuho Kaneda, Hirotada Fujii, Robert A Clark, William M. Nauseef, Katsuko Kakinuma

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Two cytosolic proteins, p47-phox and p67-phox, have been shown to be essential components of the NADPH-dependent oxidase of human neutrophils, although the specific role of each of these proteins in the multicomponent electron transport complex is undetermined. The superoxide-generating activity of this oxidase can be reproduced in a cell-free system, combining cytosol and membranes from unstimulated neutrophils in the presence of fatty acid and NADPH. In the present studies, cytosol was treated with myristic acid, arachidonic acid, or sodium dodecyl sulfate in the absence of membranes and the resultant precipitate collected by centrifugation and analyzed. Both p47-phox and p67-phox precipitated in the presence of fatty acid. However, neither FAD nor FMN was localized in the precipitates, even though substantial amounts of p47-phox and p67-phox precipitated. These results suggest that neither p47-phox nor p67-phox is a flavoprotein and that neither, therefore, is the oxidase component which accepts electrons from NADPH.

Original languageEnglish (US)
Pages (from-to)376-381
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume173
Issue number1
DOIs
StatePublished - Nov 30 1990
Externally publishedYes

Fingerprint

Flavoproteins
NADPH Oxidase
Neutrophils
NADP
Oxidoreductases
Cytosol
Precipitates
Fatty Acids
Membranes
Flavin Mononucleotide
Flavin-Adenine Dinucleotide
Cell-Free System
Centrifugation
Myristic Acid
Electron Transport
Arachidonic Acid
Superoxides
Sodium Dodecyl Sulfate
Proteins
Electrons

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Two cytosolic components of the neutrophil NADPH oxidase, P47-PHOX and P67-PHOX, are not flavoproteins. / Chiba, Tsukasa; Kaneda, Mizuho; Fujii, Hirotada; Clark, Robert A; Nauseef, William M.; Kakinuma, Katsuko.

In: Biochemical and Biophysical Research Communications, Vol. 173, No. 1, 30.11.1990, p. 376-381.

Research output: Contribution to journalArticle

Chiba, Tsukasa ; Kaneda, Mizuho ; Fujii, Hirotada ; Clark, Robert A ; Nauseef, William M. ; Kakinuma, Katsuko. / Two cytosolic components of the neutrophil NADPH oxidase, P47-PHOX and P67-PHOX, are not flavoproteins. In: Biochemical and Biophysical Research Communications. 1990 ; Vol. 173, No. 1. pp. 376-381.
@article{e01e7c5ec4d548a5a42f866b7670218f,
title = "Two cytosolic components of the neutrophil NADPH oxidase, P47-PHOX and P67-PHOX, are not flavoproteins",
abstract = "Two cytosolic proteins, p47-phox and p67-phox, have been shown to be essential components of the NADPH-dependent oxidase of human neutrophils, although the specific role of each of these proteins in the multicomponent electron transport complex is undetermined. The superoxide-generating activity of this oxidase can be reproduced in a cell-free system, combining cytosol and membranes from unstimulated neutrophils in the presence of fatty acid and NADPH. In the present studies, cytosol was treated with myristic acid, arachidonic acid, or sodium dodecyl sulfate in the absence of membranes and the resultant precipitate collected by centrifugation and analyzed. Both p47-phox and p67-phox precipitated in the presence of fatty acid. However, neither FAD nor FMN was localized in the precipitates, even though substantial amounts of p47-phox and p67-phox precipitated. These results suggest that neither p47-phox nor p67-phox is a flavoprotein and that neither, therefore, is the oxidase component which accepts electrons from NADPH.",
author = "Tsukasa Chiba and Mizuho Kaneda and Hirotada Fujii and Clark, {Robert A} and Nauseef, {William M.} and Katsuko Kakinuma",
year = "1990",
month = "11",
day = "30",
doi = "10.1016/S0006-291X(05)81068-X",
language = "English (US)",
volume = "173",
pages = "376--381",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Two cytosolic components of the neutrophil NADPH oxidase, P47-PHOX and P67-PHOX, are not flavoproteins

AU - Chiba, Tsukasa

AU - Kaneda, Mizuho

AU - Fujii, Hirotada

AU - Clark, Robert A

AU - Nauseef, William M.

AU - Kakinuma, Katsuko

PY - 1990/11/30

Y1 - 1990/11/30

N2 - Two cytosolic proteins, p47-phox and p67-phox, have been shown to be essential components of the NADPH-dependent oxidase of human neutrophils, although the specific role of each of these proteins in the multicomponent electron transport complex is undetermined. The superoxide-generating activity of this oxidase can be reproduced in a cell-free system, combining cytosol and membranes from unstimulated neutrophils in the presence of fatty acid and NADPH. In the present studies, cytosol was treated with myristic acid, arachidonic acid, or sodium dodecyl sulfate in the absence of membranes and the resultant precipitate collected by centrifugation and analyzed. Both p47-phox and p67-phox precipitated in the presence of fatty acid. However, neither FAD nor FMN was localized in the precipitates, even though substantial amounts of p47-phox and p67-phox precipitated. These results suggest that neither p47-phox nor p67-phox is a flavoprotein and that neither, therefore, is the oxidase component which accepts electrons from NADPH.

AB - Two cytosolic proteins, p47-phox and p67-phox, have been shown to be essential components of the NADPH-dependent oxidase of human neutrophils, although the specific role of each of these proteins in the multicomponent electron transport complex is undetermined. The superoxide-generating activity of this oxidase can be reproduced in a cell-free system, combining cytosol and membranes from unstimulated neutrophils in the presence of fatty acid and NADPH. In the present studies, cytosol was treated with myristic acid, arachidonic acid, or sodium dodecyl sulfate in the absence of membranes and the resultant precipitate collected by centrifugation and analyzed. Both p47-phox and p67-phox precipitated in the presence of fatty acid. However, neither FAD nor FMN was localized in the precipitates, even though substantial amounts of p47-phox and p67-phox precipitated. These results suggest that neither p47-phox nor p67-phox is a flavoprotein and that neither, therefore, is the oxidase component which accepts electrons from NADPH.

UR - http://www.scopus.com/inward/record.url?scp=0025677202&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025677202&partnerID=8YFLogxK

U2 - 10.1016/S0006-291X(05)81068-X

DO - 10.1016/S0006-291X(05)81068-X

M3 - Article

VL - 173

SP - 376

EP - 381

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -