"A 5‑lipoxygenase-specific sequence motif impedes enzyme activity and c" by Erin E. Schexnaydre, Jana Gerstmeier et al.

Faculty Publications

Title

A 5‑lipoxygenase-specific sequence motif impedes enzyme activity and confers dependence on a partner protein

Authors

Erin E. Schexnaydre, Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, USA.
Jana Gerstmeier, Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, Friedrich-Schiller-University Jena, 07743 Jena, Germany.
Ulrike Garscha, Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, Friedrich-Schiller-University Jena, 07743 Jena, Germany.
Paul M. Jordan, Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, Friedrich-Schiller-University Jena, 07743 Jena, Germany.
Oliver Werz, Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, Friedrich-Schiller-University Jena, 07743 Jena, Germany.
Marcia E. Newcomer, Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, USA. Electronic address: newcomer@lsu.edu.

Document Type

Article

Publication Date

4-1-2019

Abstract

Leukotrienes (LT) are lipid mediators of the inflammatory response that play key roles in diseases such as asthma and atherosclerosis. The precursor leukotriene A (LTA) is synthesized from arachidonic acid (AA) by 5‑lipoxygenase (5-LOX), a membrane-associated enzyme, with the help of 5‑lipoxygenase-activating protein (FLAP), a nuclear transmembrane protein. In lipoxygenases the main chain carboxylate of the C-terminus is a ligand for the non-heme iron and thus part of the catalytic center. We investigated the role of a lysine-rich sequence (KKK) 20 amino acids upstream of the C-terminus unique to 5-LOX that might displace the main-chain carboxylate in the iron coordination sphere. A 5-LOX mutant in which KKK is replaced by ENL was transfected into HEK293 cells in the absence and presence of FLAP. This mutant gave ~20-fold higher 5-LOX product levels in stimulated HEK cells relative to the wild-type 5-LOX. Co-expression of the enzymes with FLAP led to an equalization of 5-LOX products detected, with wild-type 5-LOX product levels increased and those from the mutant enzyme decreased. These data suggest that the KKK motif limits 5-LOX activity and that this attenuated activity must be compensated by the presence of FLAP as a partner protein for effective LT biosynthesis.

Publication Source (Journal or Book title)

Biochimica et biophysica acta. Molecular and cell biology of lipids

First Page

543

Last Page

551

Recommended Citation

Schexnaydre, E. E., Gerstmeier, J., Garscha, U., Jordan, P. M., Werz, O., & Newcomer, M. E. (2019). A 5‑lipoxygenase-specific sequence motif impedes enzyme activity and confers dependence on a partner protein. Biochimica et biophysica acta. Molecular and cell biology of lipids, 1864 (4), 543-551. https://doi.org/10.1016/j.bbalip.2018.09.011

This document is currently not available here.

COinS