Electrophysiological properties of heteromeric TRPV4–C1 channels
| Autor: | Xiaoqiang Yao, Yu Huang, Xin Ma, Bernd Nilius, Judy Wei-Yan Wong |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
TRPV4
Cation binding Voltage-gated ion channel Chemistry Cation permeability profile Analytical chemistry Biophysics Conductance TRPV Cation Channels Cell Biology Biochemistry Cell Line Electrophysiology Förster resonance energy transfer Cations Fluorescence Resonance Energy Transfer Homomeric TRPV4–C1 Humans Binding site Heteromeric channel |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Biomembranes. 1808(12):2789-2797 |
| ISSN: | 0005-2736 |
| DOI: | 10.1016/j.bbamem.2011.07.049 |
| Popis: | We previously reported that TRPV4 and TRPC1 can co-assemble to form heteromeric TRPV4–C1 channels [12]. In the present study, we characterized some basic electrophysiological properties of heteromeric TRPV4–C1 channels. 4α-Phorbol 12,13-didecanoate (4α-PDD, a TRPV4 agonist) activated a single channel current in HEK293 cells co-expressing TRPV4 and TRPC1. The activity of the channels was abrogated by a TRPC1-targeting blocking antibody T1E3. Conductance of the channels was ~95pS for outward currents and ~83pS for inward currents. The channels with similar conductance were also recorded in cells expressing TRPV4–C1 concatamers, in which assembled channels were expected to be mostly 2V4:2C1. Fluorescence Resonance Energy Transfer (FRET) experiments confirmed the formation of a protein complex with 2V4:2C1 stoichiometry while suggesting an unlikeliness of 3V4:1C1 or 1V4:3C1 stoichiometry. Monovalent cation permeability profiles were compared between heteromeric TRPV4–C1 and homomeric TRPV4 channels. For heteromeric TRPV4–C1 channels, their permeation profile was found to fit to Eisenman sequence VI, indicative of a strong field strength cation binding site, whereas for homomeric TRPV4 channels, their permeation profile corresponded to Eisenman sequence IV for a weak field strength binding site. Compared to homomeric TRPV4 channels, heteromeric TRPV4–C1 channels were slightly more permeable to Ca2+ and had a reduced sensitivity to extracellular Ca2+ inhibition. In summary, we found that, when TRPV4 and TRPC1 were co-expressed in HEK293 cells, the predominate assembly type was 2V4:2C1. The heteromeric TRPV4–C1 channels display distinct electrophysiological properties different from those of homomeric TRPV4 channels. |
| Databáze: | OpenAIRE |
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