Structure, mechanism, and inhibition of Hedgehog acyltransferase
| Autor: | Claire E. Coupland, Sebastian A. Andrei, T. Bertie Ansell, Loic Carrique, Pramod Kumar, Lea Sefer, Rebekka A. Schwab, Eamon F.X. Byrne, Els Pardon, Jan Steyaert, Anthony I. Magee, Thomas Lanyon-Hogg, Mark S.P. Sansom, Edward W. Tate, Christian Siebold |
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| Přispěvatelé: | Department of Bio-engineering Sciences, Structural Biology Brussels, Cancer Research UK, Biotechnology and Biological Sciences Research Council (BBSRC) |
| Rok vydání: | 2021 |
| Předmět: |
Biochemistry & Molecular Biology
STRUCTURE VALIDATION animal structures PROTEINS Protein Conformation Acylation Hedgehog acyl transferase membrane-bound O-acyltransferase Heme Sonic Hedgehog signaling Molecular Dynamics Simulation CRYO-EM Article Structure-Activity Relationship Allosteric Regulation Catalytic Domain Chlorocebus aethiops Animals Humans Hedgehog Proteins Enzyme Inhibitors PALMITOYLATION Molecular Biology 11 Medical and Health Sciences Science & Technology IDENTIFICATION REFINEMENT Palmitoyl Coenzyme A ALGORITHMS Cryoelectron Microscopy RECOGNITION Membrane Proteins small molecule inhibitor drug cryo-EM structure Cell Biology molecular dynamics simulations 06 Biological Sciences palmitoyl co enzyme A HEK293 Cells embryonic structures COS Cells VISUALIZATION integral membrane protein Life Sciences & Biomedicine Acyltransferases GENERATION Developmental Biology Signal Transduction |
| Zdroj: | Molecular Cell |
| ISSN: | 1097-4164 |
| Popis: | Summary The Sonic Hedgehog (SHH) morphogen pathway is fundamental for embryonic development and stem cell maintenance and is implicated in various cancers. A key step in signaling is transfer of a palmitate group to the SHH N terminus, catalyzed by the multi-pass transmembrane enzyme Hedgehog acyltransferase (HHAT). We present the high-resolution cryo-EM structure of HHAT bound to substrate analog palmityl-coenzyme A and a SHH-mimetic megabody, revealing a heme group bound to HHAT that is essential for HHAT function. A structure of HHAT bound to potent small-molecule inhibitor IMP-1575 revealed conformational changes in the active site that occlude substrate binding. Our multidisciplinary analysis provides a detailed view of the mechanism by which HHAT adapts the membrane environment to transfer an acyl chain across the endoplasmic reticulum membrane. This structure of a membrane-bound O-acyltransferase (MBOAT) superfamily member provides a blueprint for other protein-substrate MBOATs and a template for future drug discovery. Graphical abstract Highlights • High-resolution cryo-EM structure of HHAT in complex with a SHH-mimetic megabody • Heme is bound to HHAT Cys324 in a membrane cavity and is essential for function • Palm-CoA binds in a central HHAT cavity adjacent to the catalytic histidine • The competitive inhibitor IMP-1575 causes conformational changes in the active site HHAT is a key enzyme in the Hedgehog signaling pathway and protein-substrate member of the membrane-bound O-acyltransferase (MBOAT) superfamily. Coupland et al. report the cryo-EM structures of HHAT bound to acyl-donor substrate, megabody, and inhibitor IMP-1575, providing insight into the structure-function relationship of HHAT, mechanism, and the basis for inhibition. |
| Databáze: | OpenAIRE |
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