High rates of superoxide production in skeletal-muscle mitochondria respiring on both complex I- and complex II-linked substrates

Florian L. Muller, Yuhong Liu, Muhammad A. Abdul-Ghani, Michael S. Lustgarten, Arunabh Bhattacharya, Youngmok C. Jang, Holly Van Remmen

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

Despite the considerable interest in superoxide as a potential cause of pathology, the mechanisms of its deleterious production by mitochondria remain poorly understood. Previous studies in purified mitochondria have found that the highest rates of superoxide production are observed with succinate-driven reverse-electron transfer through complex I, although the physiological importance of this pathway is disputed because it necessitates high concentrations of succinate and is thought not to occur when NAD is in the reduced state. However, very few studies have examined the rates of superoxide production with mitochondria respiring on both NADH-linked (e.g. glutamate) and complex II-linked substrates. In the present study, we find that the rates of superoxide production (measured indirectly as H2O2) with glutamate + succinate (∼1100 pmol of H2O2· min1· mg-1) were unexpectedly much higher than with succinate (∼400pmol of H2O2·min -1·mg-1) or glutamate (∼80 pmol of H 2O2·min-1 mg-1) alone. Superoxide production with glutamate + succinate remained high even at low substrate concentrations (< 1 mM), was decreased by rotenone and was completely eliminated by FCCP (carbonyl cyanide p- trifluoromethoxyphenylhydrazone), indicating that it must in large part originate from reverse-electron transfer through complex I. Similar results were obtained when glutamate was replaced with pyruvate, α-ketoglutarate or palmitoyl carnitine. In contrast, superoxide production was consistently lowered by the addition of malate (malate + succinate ∼30 pmol of H 2O2·min-1·mg-1). We propose that the inhibitory action of malate on superoxide production can be explained by oxaloacetate inhibition of complex II. In summary, the present results indicate that reverse-electron transfer-mediated superoxide production can occur under physiologically realistic substrate conditions and suggest that oxaloacetate inhibition of complex II may be an adaptive mechanism to minimize this.

Original languageEnglish (US)
Pages (from-to)491-499
Number of pages9
JournalBiochemical Journal
Volume409
Issue number2
DOIs
StatePublished - Jan 15 2008

Fingerprint

Muscle Mitochondrion
Mitochondria
Superoxides
Muscle
Skeletal Muscle
Succinic Acid
Glutamic Acid
Substrates
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Oxaloacetic Acid
Electrons
NAD
Rotenone
Carnitine
Pathology
Pyruvic Acid

Keywords

  • Complex I
  • Electron transport chain
  • HO
  • Mitochondria
  • Oxaloacetate
  • Superoxide

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

High rates of superoxide production in skeletal-muscle mitochondria respiring on both complex I- and complex II-linked substrates. / Muller, Florian L.; Liu, Yuhong; Abdul-Ghani, Muhammad A.; Lustgarten, Michael S.; Bhattacharya, Arunabh; Jang, Youngmok C.; Van Remmen, Holly.

In: Biochemical Journal, Vol. 409, No. 2, 15.01.2008, p. 491-499.

Research output: Contribution to journalArticle

Muller, Florian L. ; Liu, Yuhong ; Abdul-Ghani, Muhammad A. ; Lustgarten, Michael S. ; Bhattacharya, Arunabh ; Jang, Youngmok C. ; Van Remmen, Holly. / High rates of superoxide production in skeletal-muscle mitochondria respiring on both complex I- and complex II-linked substrates. In: Biochemical Journal. 2008 ; Vol. 409, No. 2. pp. 491-499.
@article{76b09b5ab0e34d478e1703d3957664f1,
title = "High rates of superoxide production in skeletal-muscle mitochondria respiring on both complex I- and complex II-linked substrates",
abstract = "Despite the considerable interest in superoxide as a potential cause of pathology, the mechanisms of its deleterious production by mitochondria remain poorly understood. Previous studies in purified mitochondria have found that the highest rates of superoxide production are observed with succinate-driven reverse-electron transfer through complex I, although the physiological importance of this pathway is disputed because it necessitates high concentrations of succinate and is thought not to occur when NAD is in the reduced state. However, very few studies have examined the rates of superoxide production with mitochondria respiring on both NADH-linked (e.g. glutamate) and complex II-linked substrates. In the present study, we find that the rates of superoxide production (measured indirectly as H2O2) with glutamate + succinate (∼1100 pmol of H2O2· min1· mg-1) were unexpectedly much higher than with succinate (∼400pmol of H2O2·min -1·mg-1) or glutamate (∼80 pmol of H 2O2·min-1 mg-1) alone. Superoxide production with glutamate + succinate remained high even at low substrate concentrations (< 1 mM), was decreased by rotenone and was completely eliminated by FCCP (carbonyl cyanide p- trifluoromethoxyphenylhydrazone), indicating that it must in large part originate from reverse-electron transfer through complex I. Similar results were obtained when glutamate was replaced with pyruvate, α-ketoglutarate or palmitoyl carnitine. In contrast, superoxide production was consistently lowered by the addition of malate (malate + succinate ∼30 pmol of H 2O2·min-1·mg-1). We propose that the inhibitory action of malate on superoxide production can be explained by oxaloacetate inhibition of complex II. In summary, the present results indicate that reverse-electron transfer-mediated superoxide production can occur under physiologically realistic substrate conditions and suggest that oxaloacetate inhibition of complex II may be an adaptive mechanism to minimize this.",
keywords = "Complex I, Electron transport chain, HO, Mitochondria, Oxaloacetate, Superoxide",
author = "Muller, {Florian L.} and Yuhong Liu and Abdul-Ghani, {Muhammad A.} and Lustgarten, {Michael S.} and Arunabh Bhattacharya and Jang, {Youngmok C.} and {Van Remmen}, Holly",
year = "2008",
month = "1",
day = "15",
doi = "10.1042/BJ20071162",
language = "English (US)",
volume = "409",
pages = "491--499",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - High rates of superoxide production in skeletal-muscle mitochondria respiring on both complex I- and complex II-linked substrates

AU - Muller, Florian L.

AU - Liu, Yuhong

AU - Abdul-Ghani, Muhammad A.

AU - Lustgarten, Michael S.

AU - Bhattacharya, Arunabh

AU - Jang, Youngmok C.

AU - Van Remmen, Holly

PY - 2008/1/15

Y1 - 2008/1/15

N2 - Despite the considerable interest in superoxide as a potential cause of pathology, the mechanisms of its deleterious production by mitochondria remain poorly understood. Previous studies in purified mitochondria have found that the highest rates of superoxide production are observed with succinate-driven reverse-electron transfer through complex I, although the physiological importance of this pathway is disputed because it necessitates high concentrations of succinate and is thought not to occur when NAD is in the reduced state. However, very few studies have examined the rates of superoxide production with mitochondria respiring on both NADH-linked (e.g. glutamate) and complex II-linked substrates. In the present study, we find that the rates of superoxide production (measured indirectly as H2O2) with glutamate + succinate (∼1100 pmol of H2O2· min1· mg-1) were unexpectedly much higher than with succinate (∼400pmol of H2O2·min -1·mg-1) or glutamate (∼80 pmol of H 2O2·min-1 mg-1) alone. Superoxide production with glutamate + succinate remained high even at low substrate concentrations (< 1 mM), was decreased by rotenone and was completely eliminated by FCCP (carbonyl cyanide p- trifluoromethoxyphenylhydrazone), indicating that it must in large part originate from reverse-electron transfer through complex I. Similar results were obtained when glutamate was replaced with pyruvate, α-ketoglutarate or palmitoyl carnitine. In contrast, superoxide production was consistently lowered by the addition of malate (malate + succinate ∼30 pmol of H 2O2·min-1·mg-1). We propose that the inhibitory action of malate on superoxide production can be explained by oxaloacetate inhibition of complex II. In summary, the present results indicate that reverse-electron transfer-mediated superoxide production can occur under physiologically realistic substrate conditions and suggest that oxaloacetate inhibition of complex II may be an adaptive mechanism to minimize this.

AB - Despite the considerable interest in superoxide as a potential cause of pathology, the mechanisms of its deleterious production by mitochondria remain poorly understood. Previous studies in purified mitochondria have found that the highest rates of superoxide production are observed with succinate-driven reverse-electron transfer through complex I, although the physiological importance of this pathway is disputed because it necessitates high concentrations of succinate and is thought not to occur when NAD is in the reduced state. However, very few studies have examined the rates of superoxide production with mitochondria respiring on both NADH-linked (e.g. glutamate) and complex II-linked substrates. In the present study, we find that the rates of superoxide production (measured indirectly as H2O2) with glutamate + succinate (∼1100 pmol of H2O2· min1· mg-1) were unexpectedly much higher than with succinate (∼400pmol of H2O2·min -1·mg-1) or glutamate (∼80 pmol of H 2O2·min-1 mg-1) alone. Superoxide production with glutamate + succinate remained high even at low substrate concentrations (< 1 mM), was decreased by rotenone and was completely eliminated by FCCP (carbonyl cyanide p- trifluoromethoxyphenylhydrazone), indicating that it must in large part originate from reverse-electron transfer through complex I. Similar results were obtained when glutamate was replaced with pyruvate, α-ketoglutarate or palmitoyl carnitine. In contrast, superoxide production was consistently lowered by the addition of malate (malate + succinate ∼30 pmol of H 2O2·min-1·mg-1). We propose that the inhibitory action of malate on superoxide production can be explained by oxaloacetate inhibition of complex II. In summary, the present results indicate that reverse-electron transfer-mediated superoxide production can occur under physiologically realistic substrate conditions and suggest that oxaloacetate inhibition of complex II may be an adaptive mechanism to minimize this.

KW - Complex I

KW - Electron transport chain

KW - HO

KW - Mitochondria

KW - Oxaloacetate

KW - Superoxide

UR - http://www.scopus.com/inward/record.url?scp=38749087624&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=38749087624&partnerID=8YFLogxK

U2 - 10.1042/BJ20071162

DO - 10.1042/BJ20071162

M3 - Article

C2 - 17916065

AN - SCOPUS:38749087624

VL - 409

SP - 491

EP - 499

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -