Kinetic mechanism of linoleic acid oxidation by 5-lipoxygenase from Solatium tuberosum L

O. V. Kharchenko, A. I. Kulinichenko, I. A. Butovich

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Linoleic acid oxidation by 5-lipoxygenase from Solanum tuberosum has been studied as affected by sodium dodecylsulfate (Ds-Na). The reaction system consisted of 5-lipoxygenase and mixed micelles of linoleic acid and Lubrol PX. It contained varying amounts of the enzyme effector - Ds-Na. The enzyme showed a pronounced cooperativity, and the reaction was governed by the Hill equation with h = 3.7. On the other side, increasing amounts of Ds-Na added to the system caused a tremendous increase of enzyme activity and simultaneous decline of h, with was proportional to Ds-Na concentration. Ds-Na had dual effect on 5-lipoxygenase - there was an optimal concentration of the compound (0.34 mM Lubrol PX; 0.2 mM LA; 0.13 mM Ds-Na; pH=6.3) causing the 4-fold highest activation and h=1,6. The further increase of Ds-Na led to the enzyme inhibition. If Ds-Na was 0.5 mM, h became 1. At this point, each molecule of 5-lipoxygenase bound 3 molecules of Ds-Na and 1 molecule of linoleic acid, thus the total number of occupied binding sites was 4. A kinetic scheme of 5-lipoxygenase reaction has been proposed. It was found that the enzyme's kinetic behaviour could be explaind if assumed an existence of a special noncatalytic binding centre capable of binding several (up to 3) molecules of either substrate, or effector. Such a centre can serve as an anchoring site facilitating the enzyme binding to the surface of lipid aggregates containing insolubilised substrate molecules. Replacing linoleic acid in the binding site, Ds-Na activates the enzyme, possibly due to the much more effective translocation of 5-lipoxygenase to the surface of lipid aggregates. This mechanism can be an universal alternative to the FLAP-type regulation of 5-lipoxygenase activities.

Original languageEnglish (US)
Pages (from-to)43-44
Number of pages2
JournalUkrain'skyi Biokhimichnyi Zhurnal
Volume71
Issue number4
StatePublished - 1999

Keywords

  • 5-lipoxygenase
  • Linoleic acid
  • Regulator of 5-lipoxygenase activities
  • Sodium dodecylsulfate

ASJC Scopus subject areas

  • Biochemistry

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