Browse by author
Lookup NU author(s): Dr Sabrina MackinnonORCiD, Professor Wyatt YueORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2024 The Authors. Glycolate oxidase (HAO1) catalyses the synthesis of glyoxylate, a common metabolic intermediate that causes renal failure if accumulated. HAO1 inhibition is an emerging treatment for primary hyperoxaluria, a rare disorder of glyoxylate metabolism. Here we report the first cell-based measurement of inhibitor uptake and engagement with HAO1, by adapting the cellular thermal shift assay (CETSA) based on Nano luciferase complementation and luminescence readout. By profiling the interaction between HAO1 and four well-characterised inhibitors in intact and lysed HEK293T cells, we showed that our CETSA method differentiates between low-permeability/high-engagement and high-permeability/low-engagement ligands and is able to rank HAO1 inhibitors in line with both recombinant protein methods and previously reported indirect cellular assays. Our methodology addresses the unmet need for a robust, sensitive, and scalable cellular assay to guide HAO1 inhibitor development and, in broader terms, can be rapidly adapted for other targets to simultaneously monitor compound affinity and cellular permeability.
Author(s): Mackinnon SR, Zarganes-Tzitzikas T, Adams CJ, Brennan PE, Yue WW
Publication type: Article
Publication status: Published
Journal: Biochimie
Year: 2024
Pages: ePub ahead of Print
Online publication date: 14/08/2024
Acceptance date: 13/08/2024
Date deposited: 02/09/2024
ISSN (print): 0300-9084
ISSN (electronic): 6183-1638
Publisher: Elsevier BV
URL: https://doi.org/10.1016/j.biochi.2024.08.011
DOI: 10.1016/j.biochi.2024.08.011
PubMed id: 39151880
Altmetrics provided by Altmetric
