Structural Basis of ALDH1A2 Inhibition by Irreversible and Reversible Small Molecule Inhibitors

Yan Chen, Jin Yi Zhu, Kwon Ho Hong, David C. Mikles, Gunda I. Georg, Alex S. Goldstein, John K. Amory, Ernst Schönbrunn

Research output: Contribution to journalArticlepeer-review

36 Scopus citations


Enzymes of the ALDH1A subfamily of aldehyde dehydrogenases are crucial in regulating retinoic acid (RA) signaling and have received attention as potential drug targets. ALDH1A2 is the primary RA-synthesizing enzyme in mammalian spermatogenesis and is therefore considered a viable drug target for male contraceptive development. However, only a small number of ALDH1A2 inhibitors have been reported, and information on the structure of ALDH1A2 was limited to the NAD-liganded enzyme void of substrate or inhibitors. Herein, we describe the mechanism of action of structurally unrelated reversible and irreversible inhibitors of human ALDH1A2 using direct binding studies and X-ray crystallography. All inhibitors bind to the active sites of tetrameric ALDH1A2. Compound WIN18,446 covalently reacts with the side chain of the catalytic residue Cys320, resulting in a chiral adduct in (R) configuration. The covalent adduct directly affects the neighboring NAD molecule, which assumes a contracted conformation suboptimal for the dehydrogenase reaction. The reversible inhibitors interact predominantly through direct hydrogen bonding interactions with residues in the vicinity of Cys320 without affecting NAD. Upon interaction with inhibitors, a large flexible loop assumes regular structure, thereby shielding the active site from solvent. The precise knowledge of the binding modes provides a new framework for the rational design of novel inhibitors of ALDH1A2 with improved potency and selectivity profiles.

Original languageEnglish (US)
Pages (from-to)582-590
Number of pages9
JournalACS Chemical Biology
Issue number3
StatePublished - Mar 16 2018

Bibliographical note

Funding Information:
We thank the Moffitt Chemical Biology Core for access to the protein crystallography facility and ITC instrument (National Cancer Institute grant P30-CA076292) and the Southeast Regional Collaborative Access Team (SER-CAT, University of Georgia) for synchrotron data collection. This work was supported by the Eunice Kennedy Shriver National Institute for Child Health & Human Development (NICHD) grants HHSN275201300017C and U54HD04245.

Publisher Copyright:
© 2017 American Chemical Society.


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