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Overexpression of human transient receptor potential M5 upregulates endogenous human transient receptor potential A1 in a stable HEK cell line.
Assay Drug Dev Technol. 2010 Dec; 8(6):695-702.AD

Abstract

Transient receptor potential M5 (TRPM5), a monovalent cation channel, is primarily activated by increases in intracellular calcium. However, we found unexpectedly that allyl isothiocyanate (AITC) and structural analogs triggered a membrane potential and calcium dye responses in TRPM5-HEK cells (AITC EC₅₀ = 9.0 ± 2.4 μM, n = 5). Although AITC and its analogs were more potent on transient receptor potential A1 (TRPA1)-HEK cells (AITC EC₅₀ = 0.23 ± 0.03 μM, n = 4), the rank order potency of these compounds were similar for TRPM5- and TRPA1-HEK cells. No response to these compounds was seen in parental HEK cells, TRPM5-CHO cells, and TRPM4b-, TRPM8-, or TRPV1-transfected HEK cells. An AITC-evoked current in TRPM5-HEK cells was confirmed in whole-cell voltage clamp recording. AITC elicited an intracellular calcium increase that was not dependent on phorpholipase C(β)₂ (PLC(β)₂) activation but was dependent on extracellular calcium concentration. TRPA1 mRNA was upregulated fourfold in TRPM5-HEK cells compared with parental cells. In contrast, TRPA1 was not upregulated in HEK cells transfected in a similar manner with TRPV1 or TRPM8 genes. The AITC response was blocked by a TRPA1 inhibitor and reduced by a TRPM5 inhibitor and by targeted TRPA1 siRNA. These results suggest that TRPM5 may play a role in upregulating endogenous expression of TRPA1, that TRPA1 activation may be an additional trigger for co-expressed calcium-dependent ion channels such as TRPM5, and that TRPM5 may amplify responses to TRPA1 ligands.

Authors+Show Affiliations

Redpoint Bio Corporation, Ewing, New Jersey, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

21133676

Citation

Buber, M Tulu, et al. "Overexpression of Human Transient Receptor Potential M5 Upregulates Endogenous Human Transient Receptor Potential A1 in a Stable HEK Cell Line." Assay and Drug Development Technologies, vol. 8, no. 6, 2010, pp. 695-702.
Buber MT, Cerne R, Cortés RY, et al. Overexpression of human transient receptor potential M5 upregulates endogenous human transient receptor potential A1 in a stable HEK cell line. Assay Drug Dev Technol. 2010;8(6):695-702.
Buber, M. T., Cerne, R., Cortés, R. Y., Bryant, R. W., & Lee, S. P. (2010). Overexpression of human transient receptor potential M5 upregulates endogenous human transient receptor potential A1 in a stable HEK cell line. Assay and Drug Development Technologies, 8(6), 695-702. https://doi.org/10.1089/adt.2010.0316
Buber MT, et al. Overexpression of Human Transient Receptor Potential M5 Upregulates Endogenous Human Transient Receptor Potential A1 in a Stable HEK Cell Line. Assay Drug Dev Technol. 2010;8(6):695-702. PubMed PMID: 21133676.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Overexpression of human transient receptor potential M5 upregulates endogenous human transient receptor potential A1 in a stable HEK cell line. AU - Buber,M Tulu, AU - Cerne,Rok, AU - Cortés,Rosa Y, AU - Bryant,Robert W, AU - Lee,Seunghun Paul, Y1 - 2010/12/06/ PY - 2010/12/8/entrez PY - 2010/12/8/pubmed PY - 2011/6/24/medline SP - 695 EP - 702 JF - Assay and drug development technologies JO - Assay Drug Dev Technol VL - 8 IS - 6 N2 - Transient receptor potential M5 (TRPM5), a monovalent cation channel, is primarily activated by increases in intracellular calcium. However, we found unexpectedly that allyl isothiocyanate (AITC) and structural analogs triggered a membrane potential and calcium dye responses in TRPM5-HEK cells (AITC EC₅₀ = 9.0 ± 2.4 μM, n = 5). Although AITC and its analogs were more potent on transient receptor potential A1 (TRPA1)-HEK cells (AITC EC₅₀ = 0.23 ± 0.03 μM, n = 4), the rank order potency of these compounds were similar for TRPM5- and TRPA1-HEK cells. No response to these compounds was seen in parental HEK cells, TRPM5-CHO cells, and TRPM4b-, TRPM8-, or TRPV1-transfected HEK cells. An AITC-evoked current in TRPM5-HEK cells was confirmed in whole-cell voltage clamp recording. AITC elicited an intracellular calcium increase that was not dependent on phorpholipase C(β)₂ (PLC(β)₂) activation but was dependent on extracellular calcium concentration. TRPA1 mRNA was upregulated fourfold in TRPM5-HEK cells compared with parental cells. In contrast, TRPA1 was not upregulated in HEK cells transfected in a similar manner with TRPV1 or TRPM8 genes. The AITC response was blocked by a TRPA1 inhibitor and reduced by a TRPM5 inhibitor and by targeted TRPA1 siRNA. These results suggest that TRPM5 may play a role in upregulating endogenous expression of TRPA1, that TRPA1 activation may be an additional trigger for co-expressed calcium-dependent ion channels such as TRPM5, and that TRPM5 may amplify responses to TRPA1 ligands. SN - 1557-8127 UR - https://www.unboundmedicine.com/medline/citation/21133676/Overexpression_of_human_transient_receptor_potential_M5_upregulates_endogenous_human_transient_receptor_potential_A1_in_a_stable_HEK_cell_line_ L2 - https://www.liebertpub.com/doi/10.1089/adt.2010.0316?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -