Native chemical ligation approach to sensitively probe tissue acyl-CoA pools.
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Authors
James, Andrew M
Norman, Abigail AI
Houghton, Jack W
Prag, Hiran A
Logan, Angela
Antrobus, Robin
Hartley, Richard C
Murphy, Michael P
Publication Date
2022-07-21Journal Title
Cell Chem Biol
ISSN
2451-9456
Publisher
Elsevier BV
Type
Article
This Version
AM
Metadata
Show full item recordCitation
James, A. M., Norman, A. A., Houghton, J. W., Prag, H. A., Logan, A., Antrobus, R., Hartley, R. C., & et al. (2022). Native chemical ligation approach to sensitively probe tissue acyl-CoA pools.. Cell Chem Biol https://doi.org/10.1016/j.chembiol.2022.04.005
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.
Sponsorship
MRC (MC_UU_00015/3)
Wellcome Trust (WELL-)
Medical Research Council (MC_UU_00015/3)
Identifiers
External DOI: https://doi.org/10.1016/j.chembiol.2022.04.005
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336351
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