Electrolysis-mediated irreversible inactivation of lipoxygenase directed toward electroaffinity labelling

Thomas J. Holmes, Jonathan L. Vennerstrom, Varghese John

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Irreversible inhibition of soybean lipoxygenase-1 (SL-1) was accomplished via a controlled potential oxidative electrolysis of 1,5-dihydroxynaphthalene (1,5-DHN) at +0.8 V vs SCE. The inactivation of SL-1 with this known inhibitor was greatly enhanced under these electrolytic conditions to which the enzyme itself was stable. Electrolyses were run at 0°C in a 0.05 M phosphate buffer, pH 7.0, using graphite cloth electrodes. The rate of inactivation was observed to be limited by and dependent on the anodic oxidation of 1,5-DHN. The non-oxidizable (at this potential) inhibitor indomethacin was shown to protect the enzyme from irreversible inactivation, however, an external nucleophile (2-mercaptoethanol) had little effect. These initial studies support the capability of such electrochemical methods for the site-specific covalent modification (affinity labelling) of lipoxygenase, and perhaps other enzymes.

Original languageEnglish (US)
Pages (from-to)156-162
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume123
Issue number1
DOIs
StatePublished - Aug 30 1984

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Electrolysis
Lipoxygenase
Labeling
Enzymes
Nucleophiles
Graphite
Mercaptoethanol
Anodic oxidation
Indomethacin
Buffers
Electrodes
Phosphates
1,5-dihydroxynaphthalene
lipoxygenase L-1

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Electrolysis-mediated irreversible inactivation of lipoxygenase directed toward electroaffinity labelling. / Holmes, Thomas J.; Vennerstrom, Jonathan L.; John, Varghese.

In: Biochemical and Biophysical Research Communications, Vol. 123, No. 1, 30.08.1984, p. 156-162.

Research output: Contribution to journalArticle

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