Journal article
A Small Molecule Inhibitor of Endoplasmic Reticulum Oxidation 1 (ERO1) with Selectively Reversible Thiol Reactivity
Journal of Biological Chemistry, Vol.285(27), pp.20993-21003
Jul/2010
Abstract
Endoplasmic reticulum oxidation 1 (ERO1) is a conserved eukaryotic flavin adenine nucleotide-containing enzyme that promotes disulfide bond formation by accepting electrons from reduced protein disulfide isomerase (PDI) and passing them on to molecular oxygen. Although disulfide bond formation is an essential process, recent experiments suggest a surprisingly broad tolerance to genetic manipulations that attenuate the rate of disulfide bond formation and that a hyperoxidizing ER may place stressed cells at a disadvantage. In this study, we report on the development of a high throughput in vitro assay for mammalian ERO1 alpha activity and its application to identify small molecule inhibitors. The inhibitor EN460 (IC(50), 1.9 mu M) interacts selectively with the reduced, active form of ERO1 alpha and prevents its reoxidation. Despite rapid and promiscuous reactivity with thiolates, EN460 exhibits selectivity for ERO1. This selectivity is explained by the rapid reversibility of the reaction of EN460 with unstructured thiols, in contrast to the formation of a stable bond with ERO1 alpha followed by displacement of bound flavin adenine dinucleotide from the active site of the enzyme. Modest concentrations of EN460 and a functionally related inhibitor, QM295, promote signaling in the unfolded protein response and precondition cells against severe ER stress. Together, these observations point to the feasibility of targeting the enzymatic activity of ERO1 alpha with small molecule inhibitors.
Details
- Title
- A Small Molecule Inhibitor of Endoplasmic Reticulum Oxidation 1 (ERO1) with Selectively Reversible Thiol Reactivity
- Creators
- Jaime D. Blais (null)King-Tung Chin (null)Ester Zito (null)Yuhong Zhang (null)Nimrod Heldman (null)Heather P. Harding (null)Deborah Fass (null) - 972WIS_INST___9Colin Thorpe (null)David Ron (null)
- Resource Type
- Journal article
- Publication Details
- Journal of Biological Chemistry, Vol.285(27), pp.20993-21003; Jul/2010
- Number of pages
- 11
- Language
- English
- DOI
- https://doi.org/10.1074/jbc.M110.126599
- Grant note
- National Institutes of Health [DK075311, DK47119, ES08681, GM26643]; National Cancer Institute of Canada; Canadian Institute of Health ResearchThis work was supported, in whole or in part, by National Institutes of Health Grants DK075311, DK47119, and ES08681(to D. R.) and GM26643(to C. T.). This work was also supported by fellowships from the National Cancer Institute of Canada and the Canadian Institute of Health Research (to J.D.B.)._ALMAME_DELIMITER_
- Record Identifier
- 993264785703596
Metrics
13 Record Views