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Photochemical Probe Identification of a Small‐Molecule Inhibitor Binding Site in Hedgehog Acyltransferase (HHAT) **
Author(s) -
LanyonHogg Thomas,
Ritzefeld Markus,
Zhang Leran,
Andrei Sebastian A.,
Pogranyi Balazs,
Mondal Milon,
Sefer Lea,
Johnston Callum D.,
Coupland Claire E.,
Greenfield Jake L.,
Newington Joshua,
Fuchter Matthew J.,
Magee Anthony I.,
Siebold Christian,
Tate Edward W.
Publication year - 2021
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202014457
Subject(s) - small molecule , chemistry , binding site , hedgehog , homology modeling , stereochemistry , acyltransferase , biochemistry , enzyme , active site , hedgehog signaling pathway , biology , biophysics , computational biology , signal transduction
The mammalian membrane‐bound O ‐acyltransferase (MBOAT) superfamily is involved in biological processes including growth, development and appetite sensing. MBOATs are attractive drug targets in cancer and obesity; however, information on the binding site and molecular mechanisms underlying small‐molecule inhibition is elusive. This study reports rational development of a photochemical probe to interrogate a novel small‐molecule inhibitor binding site in the human MBOAT Hedgehog acyltransferase (HHAT). Structure‐activity relationship investigation identified single enantiomer IMP‐1575 , the most potent HHAT inhibitor reported to‐date, and guided design of photocrosslinking probes that maintained HHAT‐inhibitory potency. Photocrosslinking and proteomic sequencing of HHAT delivered identification of the first small‐molecule binding site in a mammalian MBOAT. Topology and homology data suggested a potential mechanism for HHAT inhibition which was confirmed by kinetic analysis. Our results provide an optimal HHAT tool inhibitor IMP‐1575 ( K i =38 nM) and a strategy for mapping small molecule interaction sites in MBOATs.