Photochemical probe identification of a small-molecule inhibitor binding site in hedgehog acyltransferase (HHAT)

Thomas Lanyon-Hogg; Markus Ritzefeld; Leran Zhang; Sebastian A. Andrei; Balazs Pogranyi; Milon Mondal; Lea Sefer; Callum D. Johnston; Claire E. Coupland; Jake L. Greenfield; Joshua Newington; Matthew J. Fuchter; Anthony I. Magee; Christian Siebold; Edward W. Tate

doi:10.1002/anie.202014457

Photochemical probe identification of a small-molecule inhibitor binding site in hedgehog acyltransferase (HHAT)

Thomas Lanyon-Hogg^*, Markus Ritzefeld, Leran Zhang, Sebastian A. Andrei, Balazs Pogranyi, Milon Mondal, Lea Sefer, Callum D. Johnston, Claire E. Coupland, Jake L. Greenfield, Joshua Newington, Matthew J. Fuchter, Anthony I. Magee, Christian Siebold, Edward W. Tate^*

^*Corresponding author for this work

School of Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English
Pages (from-to)	13542-13547
Number of pages	6
Journal	Angewandte Chemie International Edition
Volume	60
Issue number	24
Early online date	14 May 2021
DOIs	https://doi.org/10.1002/anie.202014457
Publication status	Published - 7 Jun 2021

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Enzymes
Hedgehog acyltransferase
Hedgehog signalling
Membrane-bound O-acyltransferase
Photoaffinity labelling

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Lanyon-Hogg, T., Ritzefeld, M., Zhang, L., Andrei, S. A., Pogranyi, B., Mondal, M., Sefer, L., Johnston, C. D., Coupland, C. E., Greenfield, J. L., Newington, J., Fuchter, M. J., Magee, A. I., Siebold, C., & Tate, E. W. (2021). Photochemical probe identification of a small-molecule inhibitor binding site in hedgehog acyltransferase (HHAT). Angewandte Chemie International Edition, 60(24), 13542-13547. https://doi.org/10.1002/anie.202014457

@article{c6361d06f266462eb7b7923df181a45a,

title = "Photochemical probe identification of a small-molecule inhibitor binding site in hedgehog acyltransferase (HHAT)",

abstract = "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.",

keywords = "Enzymes, Hedgehog acyltransferase, Hedgehog signalling, Membrane-bound O-acyltransferase, Photoaffinity labelling",

author = "Thomas Lanyon-Hogg and Markus Ritzefeld and Leran Zhang and Andrei, \{Sebastian A.\} and Balazs Pogranyi and Milon Mondal and Lea Sefer and Johnston, \{Callum D.\} and Coupland, \{Claire E.\} and Greenfield, \{Jake L.\} and Joshua Newington and Fuchter, \{Matthew J.\} and Magee, \{Anthony I.\} and Christian Siebold and Tate, \{Edward W.\}",

note = "Funding: This study was supported by Cancer Research UK grants (C6433/A16402 to A.I.M. and E.W.T, C29637/A20183 to E.W.T., and C20724/A14414 and C20724/A26752 to C.S.), the European Union Framework Program 7 (Marie Curie Intra European Fellowship to M.R. and E.W.T., and to M.M. and E.W.T.), the European Research Council under the European Union's Horizon 2020 research and innovation programme (647278 to C.S.), the Wellcome Trust (DPhil studentships 102749/Z/13/Z to L.S. and BST00110.B500.04 to C.E.C.), the BBSRC (BB/T01508X/1 to E.W.T. and C.S.), the EPSRC (EP/R00188X/1, Established Career Fellowship to M.J.F) and the Leverhulme Trust (RPG-2018-051, to J.L.G. and M.J.F.).",

year = "2021",

month = jun,

day = "7",

doi = "10.1002/anie.202014457",

language = "English",

volume = "60",

pages = "13542--13547",

journal = "Angewandte Chemie International Edition",

issn = "1433-7851",

publisher = "John Wiley \& Sons, Ltd",

number = "24",

}

Lanyon-Hogg, T, Ritzefeld, M, Zhang, L, Andrei, SA, Pogranyi, B, Mondal, M, Sefer, L, Johnston, CD, Coupland, CE, Greenfield, JL, Newington, J, Fuchter, MJ, Magee, AI, Siebold, C & Tate, EW 2021, 'Photochemical probe identification of a small-molecule inhibitor binding site in hedgehog acyltransferase (HHAT)', Angewandte Chemie International Edition, vol. 60, no. 24, pp. 13542-13547. https://doi.org/10.1002/anie.202014457

TY - JOUR

T1 - Photochemical probe identification of a small-molecule inhibitor binding site in hedgehog acyltransferase (HHAT)

AU - Lanyon-Hogg, Thomas

AU - Ritzefeld, Markus

AU - Zhang, Leran

AU - Andrei, Sebastian A.

AU - Pogranyi, Balazs

AU - Mondal, Milon

AU - Sefer, Lea

AU - Johnston, Callum D.

AU - Coupland, Claire E.

AU - Greenfield, Jake L.

AU - Newington, Joshua

AU - Fuchter, Matthew J.

AU - Magee, Anthony I.

AU - Siebold, Christian

AU - Tate, Edward W.

N1 - Funding: This study was supported by Cancer Research UK grants (C6433/A16402 to A.I.M. and E.W.T, C29637/A20183 to E.W.T., and C20724/A14414 and C20724/A26752 to C.S.), the European Union Framework Program 7 (Marie Curie Intra European Fellowship to M.R. and E.W.T., and to M.M. and E.W.T.), the European Research Council under the European Union's Horizon 2020 research and innovation programme (647278 to C.S.), the Wellcome Trust (DPhil studentships 102749/Z/13/Z to L.S. and BST00110.B500.04 to C.E.C.), the BBSRC (BB/T01508X/1 to E.W.T. and C.S.), the EPSRC (EP/R00188X/1, Established Career Fellowship to M.J.F) and the Leverhulme Trust (RPG-2018-051, to J.L.G. and M.J.F.).

PY - 2021/6/7

Y1 - 2021/6/7

N2 - 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.

AB - 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.

KW - Enzymes

KW - Hedgehog acyltransferase

KW - Hedgehog signalling

KW - Membrane-bound O-acyltransferase

KW - Photoaffinity labelling

UR - https://www.scopus.com/pages/publications/85105716926

U2 - 10.1002/anie.202014457

DO - 10.1002/anie.202014457

M3 - Article

C2 - 33768725

AN - SCOPUS:85105716926

SN - 1433-7851

VL - 60

SP - 13542

EP - 13547

JO - Angewandte Chemie International Edition

JF - Angewandte Chemie International Edition

IS - 24

ER -

Photochemical probe identification of a small-molecule inhibitor binding site in hedgehog acyltransferase (HHAT)

Abstract

UN SDGs

Keywords

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