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TRPA1 is activated by direct addition of cysteine residues to the N-hydroxysuccinyl esters of acrylic and cinnamic acids

Sadofsky, Laura; Sadofsky, Laura R.; Morice, Alyn; Boa, Andrew N.; Boa, Andrew; Maher, Sarah A.; Birrell, Mark A.; Belvisi, Maria G.; Morice, Alyn H.

Authors

Sarah A. Maher

Mark A. Birrell

Maria G. Belvisi



Abstract

The nociceptor TRPA1 is thought to be activated through covalent modification of specific cysteine residues on the N terminal of the channel. The precise mechanism of covalent modification with unsaturated carbonyl-containing compounds is unclear, therefore by examining a range of compounds which can undergo both conjugate and/or direct addition reactions we sought to further elucidate the mechanism(s) whereby TRPA1 can be activated by covalent modification. Calcium signalling was used to determine the mechanism of activation of TRPA1 expressed in HEK293 cells with a series of related compounds which were capable of either direct and/or conjugate addition processes. These results were confirmed using physiological recordings with isolated vagus nerve preparations. We found negligible channel activation with chemicals which could only react with cysteine residues via conjugate addition such as acrylamide, acrylic acid, and cinnamic acid. Compounds able to react via either conjugate or direct addition, such as acrolein, methyl vinyl ketone, mesityl oxide, acrylic acid NHS ester, cinnamaldehyde and cinnamic acid NHS ester, activated TRPA1 in a concentration dependent manner as did compounds only capable of direct addition, namely propionic acid NHS ester and hydrocinnamic acid NHS ester. These compounds failed to activate TRPV1 expressed in HEK293 cells or mock transfected HEK293 cells. For molecules capable of direct or conjugate additions, the results suggest for the first time that TRPA1 may be activated preferentially by direct addition of the thiol group of TRPA1 cysteines to the agonist carbonyl carbon of α,β-unsaturated carbonyl-containing compounds.

Citation

Sadofsky, L. R., Boa, A. N., Maher, S. A., Birrell, M. A., Belvisi, M. G., & Morice, A. H. (2011). TRPA1 is activated by direct addition of cysteine residues to the N-hydroxysuccinyl esters of acrylic and cinnamic acids. Pharmacological research : the official journal of the Italian Pharmacological Society, 63(1), 30-36. https://doi.org/10.1016/j.phrs.2010.11.004

Journal Article Type Article
Acceptance Date Nov 11, 2010
Online Publication Date Nov 19, 2010
Publication Date 2011-01
Publicly Available Date Mar 28, 2024
Journal Pharmacological Research
Print ISSN 1043-6618
Electronic ISSN 1096-1186
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 63
Issue 1
Pages 30-36
DOI https://doi.org/10.1016/j.phrs.2010.11.004
Keywords Transient receptor potential ankyrin 1; TRPA1; Cysteine covalent modification; Direct addition; N-Hydroxysuccinyl esters
Public URL https://hull-repository.worktribe.com/output/424074
Publisher URL https://www.sciencedirect.com/science/article/abs/pii/S1043661810002173?via%3Dihub