Cho, Chang-Hoon Kim, Eunju Lee, Young-Sun Yarishkin, Oleg Yoo, Jae Cheal Park, Jae-Yong Hong, Seong-Geun Hwang, Eun Mi 2024-01-20T09:30:50Z 2024-01-20T09:30:50Z 2022-01-10 2014-07-22 1756-6606 https://pubs.kist.re.kr/handle/201004/126580 Background: TRPM4 channels are Ca2+ activated nonselective cation channels which are deeply involved in physiological and pathological conditions. However, their trafficking mechanism and binding partners are still elusive. Results: We have found the 14-3-3 gamma as a binding partner for TRPM4b using its N-terminal fragment from the yeast-two hybrid screening. Ser88 at the N-terminus of TRPM4b is critical for 14-3-3 gamma binding by showing GST pull-down and co-immunoprecipitation. Heterologous overexpression of 14-3-3 gamma in HEK293T cells increased TRPM4b expression on the plasma membrane which was measured by whole-cell recordings and cell surface biotinylation experiment. Surface expression of TRPM4b was greatly reduced by short hairpin RNA (shRNA) against 14-3-3 gamma. Next, endogenous TRPM4b-mediated currents were electrophysiologically characterized by application of glutamate and 9-phenanthrol, a TRPM4b specific antagonist in HT-22 cells which originated from mouse hippocampal neurons. Glutamate-induced TRPM4b currents were significantly attenuated by shRNAs against 14-3-3 gamma or TRPM4b in these cells. Finally, glutamate-induced cell death was greatly prevented by treatment of 9-phenanthrol or 14-3-3 gamma shRNA. Conclusion: These results showed that the cell surface expression of TRPM4 channels is mediated by 14-3-3 gamma binding, and the specific inhibition of this trafficking process can be a potential therapeutic target for glutamate-induced neuronal cell death. English BMC CATION CHANNEL INTRACELLULAR CALCIUM KAINATE RECEPTORS SINGLE-CHANNEL PROTEINS 14-3-3-PROTEINS TRAFFICKING SENSITIVITY MODULATION ISOFORMS Depletion of 14-3-3 gamma reduces the surface expression of Transient Receptor Potential Melastatin 4b (TRPM4b) Channels and attenuates TRPM4b-mediated glutamate-induced neuronal cell death Article 10.1186/s13041-014-0052-3 1 MOLECULAR BRAIN, v.7 MOLECULAR BRAIN 7 scie scopus 000339587900001 2-s2.0-84904775131 Neurosciences Neurosciences & Neurology Article CATION CHANNEL INTRACELLULAR CALCIUM KAINATE RECEPTORS SINGLE-CHANNEL PROTEINS 14-3-3-PROTEINS TRAFFICKING SENSITIVITY MODULATION ISOFORMS TRPM4b 14-3-3 Non-selective cation channels 9-phenanthrol HT-22 Protein-protein interaction Surface expression Hippocampal neurons Calcium activated cation channels MTT assay