Structural and functional evaluation mammalian and plant lipoxygenases upon association with nanodics as membrane mimetics

Sinem Ulusan, Ilir Sheraj, Sabine Stehling, Igor Ivanov, Aditi Das, Hartmut Kühn, Sreeparna Banerjee

Research output: Contribution to journalArticlepeer-review

Abstract

Lipoxygenases (LOX) are a family lipid oxygenating enzymes that can generate bioactive lipids of clinical relevance from polyunsaturated fatty acids. Most LOXs display a Ca2+-dependent association with membranes for their activity. Nanodiscs (ND) are stable self-assembled discoidal fragments of lipid bilayers that can mimic the plasma membrane. In this study, we evaluated the association of mammalian 15-LOXs (ALOX15 and ALOX15B) and soybean LOX-1 with NDs (LOX-ND), their enzymatic activities and inhibition. Mammalian LOXs associated with NDs showed better retention of enzymatic function compared to soybean LOX-1. Treatment of both LOX-NDs and free enzymes with the pan-LOX inhibitor nordihydroguaiaretic acid (NDGA) showed an approximately 5-fold more effective inhibition of the enzymes associated with NDs compared to the free form. NDs are easy to generate membrane mimics that can be used as an effective tool to determine enzymatic function and inhibition of membrane associated proteins.

Original languageEnglish (US)
Article number106855
JournalBiophysical Chemistry
Volume288
DOIs
StatePublished - Sep 2022

Keywords

  • Enzyme activity
  • Inhibition
  • Lipoxygenase
  • Nanodisc

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Organic Chemistry

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