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PMID: 6288006 Published · ppublish English Journal Article

A quantitative test for superoxide radicals produced in biological systems.

The Biochemical journal ·Vol. 203 ·No. 3 ·1982-06-01 ·Pages 551-8

Kuthan H, Ullrich V, Estabrook RW

Abstract

The preparation and properties of a partially succinoylated cytochrome c, suited for the detection of superoxide anion radicals in liver microsomes, is reported. By succinoylation of 45% of the primary amino groups of horse heart cytochrome c the activity towards solubilized NADPH--cytochrome P-450 reductase was diminished by 99% compared with native cytochrome c. The capacities of cytochrome b5 and cytochrome c oxidase to reduce the succinoylated ferricytochrome c and oxidize succinoylated ferrocytochrome c respectively were decreased to a similar extent. However, the bimolecular rate constant for the reduction of the partially succinoylated ferricytochrome c by O2-. was estimated to be one-tenth of the value for the reaction of O2-. with native ferricytochrome c a pH 7.7. On this basis the quantification of O2-. generated by NADPH-supplemented liver microsomes became possible. The initial rates of succinoylated ferricytochrome c reduction determined at various finite concentrations of the cytochrome c derivative can be extrapolated to obtain true rates of O2-. generation in a homogeneous system. The problems encountered in the quantitative determination of O2-. produced in biological membranes, e.g. microsomes, are discussed.

MeSH Terms
Animals Cytochrome c Group/metabolism Free Radicals In Vitro Techniques Kinetics Male Methods Microsomes, Liver/analysis Oxidation-Reduction Oxygen/analysis Rats Rats, Inbred Strains Superoxide Dismutase/metabolism Superoxides/analysis Xanthine Oxidase/metabolism
Chemicals
Cytochrome c Group Free Radicals Superoxides Superoxide Dismutase Xanthine Oxidase Oxygen
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kuthan H
Ullrich V
Estabrook R W
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32 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2030551
Published
1982-06-01
Pages
551-8
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1158268
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2030551 PMC Full Text View journal details
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