Photochemical Probe Identification of a Small-Molecule Inhibitor Binding Site in Hedgehog Acyltransferase (HHAT)*.
Thomas Lanyon-HoggMarkus RitzefeldLeran ZhangSebastian A AndreiBalazs PogranyiMilon MondalLea SeferCallum D JohnstonClaire E CouplandJake L GreenfieldJoshua NewingtonMatthew John FuchterAnthony I MageeChristian SieboldEdward William TatePublished in: Angewandte Chemie (International ed. in English) (2021)
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 (Ki =38 nM) and a strategy for mapping small molecule interaction sites in MBOATs.
Keyphrases
- small molecule
- protein protein
- endothelial cells
- structure activity relationship
- metabolic syndrome
- living cells
- emergency department
- high resolution
- machine learning
- electronic health record
- risk assessment
- papillary thyroid
- physical activity
- climate change
- deep learning
- artificial intelligence
- human health
- lymph node