Electrophile-Induced Conformational Switch of the Human TRPA1 Ion Channel Detected by Mass Spectrometry

Moparthi, Lavanya; Kjellström, Sven; Kjellbom, Per; Filipovic, Milos R.; Zygmunt, Peter M.; Johanson, Urban

doi:10.3390/ijms21186667

Open AccessArticle

Electrophile-Induced Conformational Switch of the Human TRPA1 Ion Channel Detected by Mass Spectrometry

by

Lavanya Moparthi

^1,2,3

,

Sven Kjellström

⁴,

Per Kjellbom

³,

Milos R. Filipovic

^5,6,

Peter M. Zygmunt

^7,*

and

Urban Johanson

^3,*

¹

Wallenberg Centre for Molecular Medicine (WCMM), Linköping University, SE-581 85 Linköping, Sweden

²

Department of Biomedical and Clinical Sciences (BKV), Faculty of Health Sciences, Linköping University, SE-581 85 Linköping, Sweden

³

Division of Biochemistry and Structural Biology, Center for Molecular Protein Science, Lund University, PO Box 124, SE-221 00 Lund, Sweden

⁴

Division of Mass Spectrometry, Department of Clinical Sciences, Lund University, SE-22184 Lund, Sweden

⁵

Department of Chemistry and Pharmacy, Friedrich-Alexander University Erlangen-Nuremberg, Egerlandstrasse 1, 91058 Erlangen, Germany

⁶

IBGC, UMR 5095, Universite de Bordeaux, 1, rue Camille Saint Saëns, CS 61390, 33077 Bordeaux CEDEX, France

⁷

Department of Clinical Sciences Malmö, Lund University, SE-214 28 Malmö, Sweden

^*

Authors to whom correspondence should be addressed.

Int. J. Mol. Sci. 2020, 21(18), 6667; https://doi.org/10.3390/ijms21186667

Submission received: 6 August 2020 / Revised: 6 September 2020 / Accepted: 9 September 2020 / Published: 11 September 2020

(This article belongs to the Special Issue TRPA1 Channel)

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Abstract

The human Transient Receptor Potential A1 (hTRPA1) ion channel, also known as the wasabi receptor, acts as a biosensor of various potentially harmful stimuli. It is activated by a wide range of chemicals, including the electrophilic compound N-methylmaleimide (NMM), but the mechanism of activation is not fully understood. Here, we used mass spectrometry to map and quantify the covalent labeling in hTRPA1 at three different concentrations of NMM. A functional truncated version of hTRPA1 (Δ1-688 hTRPA1), lacking the large N-terminal ankyrin repeat domain (ARD), was also assessed in the same way. In the full length hTRPA1, the labeling of different cysteines ranged from nil up to 95% already at the lowest concentration of NMM, suggesting large differences in reactivity of the thiols. Most important, the labeling of some cysteine residues increased while others decreased with the concentration of NMM, both in the full length and the truncated protein. These findings indicate a conformational switch of the proteins, possibly associated with activation or desensitization of the ion channel. In addition, several lysines in the transmembrane domain and the proximal N-terminal region were labeled by NMM, raising the possibility that lysines are also key targets for electrophilic activation of hTRPA1.

Keywords: TRP channel; TRPA1; wasabi receptor; electrophile sensing; N-methylmaleimide; iodoacetamide; alkylated thiols; redox sensitivity; pain; mass spectrometry

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MDPI and ACS Style

Moparthi, L.; Kjellström, S.; Kjellbom, P.; Filipovic, M.R.; Zygmunt, P.M.; Johanson, U. Electrophile-Induced Conformational Switch of the Human TRPA1 Ion Channel Detected by Mass Spectrometry. Int. J. Mol. Sci. 2020, 21, 6667. https://doi.org/10.3390/ijms21186667

AMA Style

Moparthi L, Kjellström S, Kjellbom P, Filipovic MR, Zygmunt PM, Johanson U. Electrophile-Induced Conformational Switch of the Human TRPA1 Ion Channel Detected by Mass Spectrometry. International Journal of Molecular Sciences. 2020; 21(18):6667. https://doi.org/10.3390/ijms21186667

Chicago/Turabian Style

Moparthi, Lavanya, Sven Kjellström, Per Kjellbom, Milos R. Filipovic, Peter M. Zygmunt, and Urban Johanson. 2020. "Electrophile-Induced Conformational Switch of the Human TRPA1 Ion Channel Detected by Mass Spectrometry" International Journal of Molecular Sciences 21, no. 18: 6667. https://doi.org/10.3390/ijms21186667

APA Style

Moparthi, L., Kjellström, S., Kjellbom, P., Filipovic, M. R., Zygmunt, P. M., & Johanson, U. (2020). Electrophile-Induced Conformational Switch of the Human TRPA1 Ion Channel Detected by Mass Spectrometry. International Journal of Molecular Sciences, 21(18), 6667. https://doi.org/10.3390/ijms21186667

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Electrophile-Induced Conformational Switch of the Human TRPA1 Ion Channel Detected by Mass Spectrometry

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