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PMID: 6439185 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Composition of partially purified NADPH oxidase from pig neutrophils.

The Biochemical journal ·Vol. 223 ·No. 3 ·1984-11-01 ·Pages 639-48

Bellavite P, Jones OT, Cross AR, Papini E, Rossi F

Abstract

The superoxide (O2.-)-forming enzyme NADPH oxidase from pig neutrophils was solubilized and partially purified by gel-filtration chromatography. The purification procedure allowed the separation of NADPH oxidase activity from NADH-dependent cytochrome c reductase and 2,6-dichlorophenol-indophenol reductase activities. O2.-forming activity was co-purified with cytochrome b-245 and was associated with phospholipids. However, active fractions endowed with cytochrome b were devoid of ubiquinone and contained only little FAD. The cytochrome b/FAD ratio was 1.13:1 in the crude solubilized extract and increased to 18.95:1 in the partially purified preparations. Most of FAD was associated with fractions containing NADH-dependent oxidoreductases. These results are consistent with the postulated role of cytochrome b in O2.-formation by neutrophil NADPH oxidase, but raise doubts about the participation of flavoproteins in this enzyme activity.

MeSH Terms
Animals Chromatography, Gel Chromatography, High Pressure Liquid Cytochrome b Group/blood Flavin-Adenine Dinucleotide/blood NADH, NADPH Oxidoreductases/blood,isolation & purification NADPH Oxidases NADPH-Ferrihemoprotein Reductase/blood Neutrophils/enzymology Quinone Reductases/blood Subcellular Fractions/enzymology Swine Ubiquinone/blood
Chemicals
Cytochrome b Group Ubiquinone Flavin-Adenine Dinucleotide NADH, NADPH Oxidoreductases NADPH-Ferrihemoprotein Reductase NADPH Oxidases Quinone Reductases dichlorophenolindophenol reductase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Bellavite P
Jones O T
Cross A R
Papini E
Rossi F
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34 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2230639
Published
1984-11-01
Pages
639-48
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1144347
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2230639 PMC Full Text View journal details
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