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Cells 2018, 7(9), 126; https://doi.org/10.3390/cells7090126

Transient Receptor Potential Channel A1 (TRPA1) Regulates Sulfur Mustard-Induced Expression of Heat Shock 70 kDa Protein 6 (HSPA6) In Vitro

1
Bundeswehr Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universität Munich, 80937 Munich, Germany
2
Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universität Munich, 80336 Munich, Germany
*
Author to whom correspondence should be addressed.
Received: 26 July 2018 / Revised: 23 August 2018 / Accepted: 28 August 2018 / Published: 31 August 2018
(This article belongs to the Special Issue TRP Channels in Health and Disease)
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Abstract

The chemosensory transient receptor potential ankyrin 1 (TRPA1) ion channel perceives different sensory stimuli. It also interacts with reactive exogenous compounds including the chemical warfare agent sulfur mustard (SM). Activation of TRPA1 by SM results in elevation of intracellular calcium levels but the cellular consequences are not understood so far. In the present study we analyzed SM-induced and TRPA1-mediated effects in human TRPA1-overexpressing HEK cells (HEKA1) and human lung epithelial cells (A549) that endogenously exhibit TRPA1. The specific TRPA1 inhibitor AP18 was used to distinguish between SM-induced and TRPA1-mediated or TRPA1-independent effects. Cells were exposed to 600 µM SM and proteome changes were investigated 24 h afterwards by 2D gel electrophoresis. Protein spots with differential staining levels were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nano liquid chromatography electrospray ionization tandem mass spectrometry. Results were verified by RT-qPCR experiments in both HEKA1 or A549 cells. Heat shock 70 kDa protein 6 (HSPA6) was identified as an SM-induced and TRPA1-mediated protein. AP18 pre-treatment diminished the up-regulation. RT-qPCR measurements verified these results and further revealed a time-dependent regulation. Our results demonstrate that SM-mediated activation of TRPA1 influences the protein expression and confirm the important role of TRPA1 ion channels in the molecular toxicology of SM. View Full-Text
Keywords: 2D gel electrophoresis; AP18; HEK293; HSP70; MALDI-TOF MS(/MS); nanoHPLC-ESI MS/MS; proteomics; sulfur mustard; TRPA1 2D gel electrophoresis; AP18; HEK293; HSP70; MALDI-TOF MS(/MS); nanoHPLC-ESI MS/MS; proteomics; sulfur mustard; TRPA1
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Lüling, R.; John, H.; Gudermann, T.; Thiermann, H.; Mückter, H.; Popp, T.; Steinritz, D. Transient Receptor Potential Channel A1 (TRPA1) Regulates Sulfur Mustard-Induced Expression of Heat Shock 70 kDa Protein 6 (HSPA6) In Vitro. Cells 2018, 7, 126.

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