Structure and Mechanism of Hedgehog Acyl Transferase
SUMMARY The iconic Sonic Hedgehog (SHH) morphogen pathway is a fundamental orchestrator of embryonic development and stem cell maintenance, and is implicated in cancers in various organs. A key step in signalling is transfer of a palmitate group to the N-terminal cysteine residue of SHH, catalysed by the multi-pass transmembrane enzyme Hedgehog acyltransferase (HHAT) resident in the endoplasmic reticulum (ER). Here, we present the high-resolution cryo-EM structure of HHAT bound to substrate analogue palmityl-coenzyme A and a SHH mimetic megabody. Surprisingly, we identified a heme group bound to an HHAT cysteine residue and show that this modification is essential for HHAT structure and function. A structure of HHAT bound to potent small molecule inhibitor IMP-1575 revealed conformational changes in the active site which occlude substrate binding. Our multidisciplinary analysis provides a detailed view of the novel mechanism by which HHAT adapts the membrane environment to transfer a long chain fatty acid across the ER membrane from cytosolic acyl-CoA to a luminal protein substrate. This structure of a member of the protein-substrate membrane-bound O-acyltransferase (MBOAT) superfamily provides a blueprint for other protein substrate MBOATs, such as WNT morphogen acyltransferase Porcupine and ghrelin O-acyltransferase GOAT, and a template for future drug discovery..
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Preprint| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
bioRxiv.org - (2021) vom: 12. Juli Zur Gesamtaufnahme - year:2021 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Coupland, Claire E. [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2021.07.08.451580 |
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| PPN (Katalog-ID): |
XBI032166842 |
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| 520 | |a SUMMARY The iconic Sonic Hedgehog (SHH) morphogen pathway is a fundamental orchestrator of embryonic development and stem cell maintenance, and is implicated in cancers in various organs. A key step in signalling is transfer of a palmitate group to the N-terminal cysteine residue of SHH, catalysed by the multi-pass transmembrane enzyme Hedgehog acyltransferase (HHAT) resident in the endoplasmic reticulum (ER). Here, we present the high-resolution cryo-EM structure of HHAT bound to substrate analogue palmityl-coenzyme A and a SHH mimetic megabody. Surprisingly, we identified a heme group bound to an HHAT cysteine residue and show that this modification is essential for HHAT structure and function. A structure of HHAT bound to potent small molecule inhibitor IMP-1575 revealed conformational changes in the active site which occlude substrate binding. Our multidisciplinary analysis provides a detailed view of the novel mechanism by which HHAT adapts the membrane environment to transfer a long chain fatty acid across the ER membrane from cytosolic acyl-CoA to a luminal protein substrate. This structure of a member of the protein-substrate membrane-bound O-acyltransferase (MBOAT) superfamily provides a blueprint for other protein substrate MBOATs, such as WNT morphogen acyltransferase Porcupine and ghrelin O-acyltransferase GOAT, and a template for future drug discovery. | ||
| 700 | 1 | |a Andrei, Sebastian A. |e verfasserin |4 aut | |
| 700 | 1 | |a Ansell, T. Bertie |e verfasserin |4 aut | |
| 700 | 1 | |a Carrique, Loic |e verfasserin |4 aut | |
| 700 | 1 | |a Kumar, Pramod |e verfasserin |4 aut | |
| 700 | 1 | |a Sefer, Lea |e verfasserin |4 aut | |
| 700 | 1 | |a Schwab, Rebekka A |e verfasserin |4 aut | |
| 700 | 1 | |a Byrne, Eamon F.X. |e verfasserin |4 aut | |
| 700 | 1 | |a Pardon, Els |e verfasserin |4 aut | |
| 700 | 1 | |a Steyaert, Jan |e verfasserin |4 aut | |
| 700 | 1 | |a Magee, Anthony I. |e verfasserin |4 aut | |
| 700 | 1 | |a Lanyon-Hogg, Thomas. |e verfasserin |4 aut | |
| 700 | 1 | |a Sansom, Mark S. P. |e verfasserin |4 aut | |
| 700 | 1 | |a Tate, Edward W. |e verfasserin |4 aut | |
| 700 | 1 | |a Siebold, Christian |e verfasserin |4 aut | |
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