Effect of 4-hydroxy-2(E)-nonenal on soybean lipoxygenase-1

Harold W. Gardner, Nigel Deighton

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The oxidation of linoleic acid by soybean lipoxygenase-1 (LOX-1) was inhibited in a time-dependent manner by 4-hydroxy-2(E)-nonenal (HNE). Kinetic analysis indicated the effect was due to slow-binding inhibition conforming to an affinity labeling mechanism-based inhibition. After 25 min of preincubation of LOX-1 with and without HNE, Lineweaver-Burk reciprocal plots indicated mixed noncompetitive/competitive inhibition. Low concentrations of HNE influenced the electron paramagnetic resonance (EPR) signal of 13(S)-hydroperoxy-9(Z), 11(E)-octadecadienoic acid (13-HPODE)-generated Fe3+-LOX-1 slightly, but higher concentrations completely eliminated the EPR signal indicating an active site hindered from access by 13-HPODE. HNE may compete for the active site of LOX-1 because its precursor, 4-hydroperoxy-(2E)-nonenal, is a product of LOX-1 oxidation of (3Z)-nonenal. Also, it was an attractive hypothesis to suggest that HNE may disrupt the active site by forming a Michael adduct with one or more of the three histidines that ligate the iron active site of LOX-1.

Original languageEnglish (US)
Article number766
Pages (from-to)623-628
Number of pages6
JournalLipids
Volume36
Issue number6
DOIs
StatePublished - Jan 1 2001

Fingerprint

Lipoxygenase
Catalytic Domain
Electron Spin Resonance Spectroscopy
Paramagnetic resonance
Oxidation
Linoleic Acid
Histidine
Labeling
Iron
4-hydroxy-2-nonenal
2-nonenal
lipoxygenase L-1
Kinetics

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Organic Chemistry
  • Cell Biology

Cite this

Gardner, Harold W. ; Deighton, Nigel. / Effect of 4-hydroxy-2(E)-nonenal on soybean lipoxygenase-1. In: Lipids. 2001 ; Vol. 36, No. 6. pp. 623-628.
@article{984c0a4a64dd43aaa0594cf72654c3c2,
title = "Effect of 4-hydroxy-2(E)-nonenal on soybean lipoxygenase-1",
abstract = "The oxidation of linoleic acid by soybean lipoxygenase-1 (LOX-1) was inhibited in a time-dependent manner by 4-hydroxy-2(E)-nonenal (HNE). Kinetic analysis indicated the effect was due to slow-binding inhibition conforming to an affinity labeling mechanism-based inhibition. After 25 min of preincubation of LOX-1 with and without HNE, Lineweaver-Burk reciprocal plots indicated mixed noncompetitive/competitive inhibition. Low concentrations of HNE influenced the electron paramagnetic resonance (EPR) signal of 13(S)-hydroperoxy-9(Z), 11(E)-octadecadienoic acid (13-HPODE)-generated Fe3+-LOX-1 slightly, but higher concentrations completely eliminated the EPR signal indicating an active site hindered from access by 13-HPODE. HNE may compete for the active site of LOX-1 because its precursor, 4-hydroperoxy-(2E)-nonenal, is a product of LOX-1 oxidation of (3Z)-nonenal. Also, it was an attractive hypothesis to suggest that HNE may disrupt the active site by forming a Michael adduct with one or more of the three histidines that ligate the iron active site of LOX-1.",
author = "Gardner, {Harold W.} and Nigel Deighton",
year = "2001",
month = "1",
day = "1",
doi = "10.1007/s11745-001-0766-9",
language = "English (US)",
volume = "36",
pages = "623--628",
journal = "Lipids",
issn = "0024-4201",
publisher = "Springer Verlag",
number = "6",

}

Effect of 4-hydroxy-2(E)-nonenal on soybean lipoxygenase-1. / Gardner, Harold W.; Deighton, Nigel.

In: Lipids, Vol. 36, No. 6, 766, 01.01.2001, p. 623-628.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effect of 4-hydroxy-2(E)-nonenal on soybean lipoxygenase-1

AU - Gardner, Harold W.

AU - Deighton, Nigel

PY - 2001/1/1

Y1 - 2001/1/1

N2 - The oxidation of linoleic acid by soybean lipoxygenase-1 (LOX-1) was inhibited in a time-dependent manner by 4-hydroxy-2(E)-nonenal (HNE). Kinetic analysis indicated the effect was due to slow-binding inhibition conforming to an affinity labeling mechanism-based inhibition. After 25 min of preincubation of LOX-1 with and without HNE, Lineweaver-Burk reciprocal plots indicated mixed noncompetitive/competitive inhibition. Low concentrations of HNE influenced the electron paramagnetic resonance (EPR) signal of 13(S)-hydroperoxy-9(Z), 11(E)-octadecadienoic acid (13-HPODE)-generated Fe3+-LOX-1 slightly, but higher concentrations completely eliminated the EPR signal indicating an active site hindered from access by 13-HPODE. HNE may compete for the active site of LOX-1 because its precursor, 4-hydroperoxy-(2E)-nonenal, is a product of LOX-1 oxidation of (3Z)-nonenal. Also, it was an attractive hypothesis to suggest that HNE may disrupt the active site by forming a Michael adduct with one or more of the three histidines that ligate the iron active site of LOX-1.

AB - The oxidation of linoleic acid by soybean lipoxygenase-1 (LOX-1) was inhibited in a time-dependent manner by 4-hydroxy-2(E)-nonenal (HNE). Kinetic analysis indicated the effect was due to slow-binding inhibition conforming to an affinity labeling mechanism-based inhibition. After 25 min of preincubation of LOX-1 with and without HNE, Lineweaver-Burk reciprocal plots indicated mixed noncompetitive/competitive inhibition. Low concentrations of HNE influenced the electron paramagnetic resonance (EPR) signal of 13(S)-hydroperoxy-9(Z), 11(E)-octadecadienoic acid (13-HPODE)-generated Fe3+-LOX-1 slightly, but higher concentrations completely eliminated the EPR signal indicating an active site hindered from access by 13-HPODE. HNE may compete for the active site of LOX-1 because its precursor, 4-hydroperoxy-(2E)-nonenal, is a product of LOX-1 oxidation of (3Z)-nonenal. Also, it was an attractive hypothesis to suggest that HNE may disrupt the active site by forming a Michael adduct with one or more of the three histidines that ligate the iron active site of LOX-1.

UR - http://www.scopus.com/inward/record.url?scp=0034904237&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034904237&partnerID=8YFLogxK

U2 - 10.1007/s11745-001-0766-9

DO - 10.1007/s11745-001-0766-9

M3 - Article

VL - 36

SP - 623

EP - 628

JO - Lipids

JF - Lipids

SN - 0024-4201

IS - 6

M1 - 766

ER -