Native chemical ligation approach to sensitively probe tissue acyl-CoA pools

James, A. M., Norman, A. A. I., Houghton, J. W., Prag, H. A., Logan, A., Antrobus, R., Hartley, R. C. and Murphy, M. P. (2022) Native chemical ligation approach to sensitively probe tissue acyl-CoA pools. Cell Chemical Biology, 29(7), 1232-1244.e5. (doi: 10.1016/j.chembiol.2022.04.005) (PMID:35868236) (PMCID:PMC9586882)

Abstract

During metabolism, carboxylic acids are often activated by conjugation to the thiol of coenzyme A (CoA). The resulting acyl-CoAs comprise a group of ∼100 thioester-containing metabolites that could modify protein behavior through non-enzymatic N-acylation of lysine residues. However, the importance of many potential acyl modifications remains unclear because antibody-based methods to detect them are unavailable and the in vivo concentrations of their respective acyl-CoAs are poorly characterized. Here, we develop cysteine-triphenylphosphonium (CysTPP), a mass spectrometry probe that uses “native chemical ligation” to sensitively detect the major acyl-CoAs present in vivo through irreversible modification of its amine via a thioester intermediate. Using CysTPP, we show that longer-chain (C13–C22) acyl-CoAs often constitute ∼60% of the acyl-CoA pool in rat tissues. These hydrophobic longer-chain fatty acyl-CoAs have the potential to non-enzymatically modify protein residues.

• Item Type: Articles
• Additional Information: This work was supported by a grant from the Medical Research Council UK to MPM (MC_UU_00015/3) and a studentship to AAIN from University of Glasgow. This research was funded in part by the Wellcome Trust (MPM: 220257/Z/20/Z and RCH: 110158/Z/15/Z). The author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Hartley, Professor Richard and Norman, Miss Abigail
• Authors: James, A. M., Norman, A. A. I., Houghton, J. W., Prag, H. A., Logan, A., Antrobus, R., Hartley, R. C., and Murphy, M. P.
• College/School: College of Science and Engineering > School of Chemistry
• Journal Name: Cell Chemical Biology
• Publisher: Elsevier (Cell Press)
• ISSN: 2451-9456
• ISSN (Online): 2451-9448
• Copyright Holders: Copyright © 2022 The Authors
• First Published: First published in Cell Chemical Biology 29(7): 1232-1244.e5
• Publisher Policy: Reproduced under a Creative Commons License
• Data DOI: 10.5525/gla.researchdata.1214

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
172617		Exploring mitochondrial metabolism in health and disease using targeted biological chemistry	Richard Hartley	Wellcome Trust (WELLCOTR)	110158/Z/15/Z	Chemistry