論文 - 詳細
| RRC ID | 83213 |
|---|---|
| 著者 | Kuwashima Y, Yanagawa M, Maekawa M, Abe M, Sako Y, Arita M. |
| タイトル | TRPV4-dependent Ca2+ influx determines cholesterol dynamics at the plasma membrane. |
| ジャーナル | Biophys J |
| Abstract |
The activities of the transient receptor potential vanilloid 4 (TRPV4), a Ca2+-permeable nonselective cation channel, are controlled by its surrounding membrane lipids (e.g., cholesterol, phosphoinositides). The transmembrane region of TRPV4 contains a cholesterol recognition amino acid consensus (CRAC) motif and its inverted (CARC) motif located in the plasmalemmal cytosolic leaflet. TRPV4 localizes in caveolae, a bulb-shaped cholesterol-rich domain at the plasma membrane. Here, we visualized the spatiotemporal interactions between TRPV4 and cholesterol at the plasma membrane in living cells by dual-color single-molecule imaging using total internal reflection fluorescence microscopy. To this aim, we labeled cholesterol at the cytosolic leaflets of the plasma membrane using a cholesterol biosensor, D4H. Our single-molecule tracking analysis showed that the TRPV4 molecules colocalize with D4H-accessible cholesterol molecules mainly in the low fluidity membrane domains in which both molecules are highly clustered. Colocalization of TRPV4 and D4H-accessible cholesterol was observed both inside and outside of caveolae. Agonist-evoked TRPV4 activation remarkably decreased colocalization probability and association rate between TRPV4 and D4H-accessible cholesterol molecules. Interestingly, upon TRPV4 activation, the particle density of D4H-accessible cholesterol molecules was decreased and the D4H-accessible cholesterol molecules in the fast-diffusing state were increased at the plasma membrane. The introduction of skeletal dysplasia-associated R616Q mutation into the CRAC/CARC motif of TRPV4, which reduced the interaction with cholesterol clusters, could not alter the D4H-accessible cholesterol dynamics. Mechanistically, TRPV4-mediated Ca2+ influx and the C-terminal calmodulin-binding site of TRPV4 are essential for modulating the plasmalemmal D4H-accessible cholesterol dynamics. We propose that TRPV4 remodels its surrounding plasmalemmal environment by manipulating cholesterol dynamics through Ca2+ influx. |
| 巻・号 | 123(7) |
| ページ | 867-884 |
| 公開日 | 2024-4-2 |
| DOI | 10.1016/j.bpj.2024.02.030 |
| PII | S0006-3495(24)00164-4 |
| PMID | 38433447 |
| PMC | PMC10995426 |
| MeSH | Calcium Signaling* Calmodulin / metabolism Cell Membrane / metabolism Cholesterol / metabolism TRPV Cation Channels* / metabolism |
| IF | 3.854 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 5 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 遺伝子材料 | pFN21A-TRPV4 (WT) (RDB20676) pFN21A HaloTag-TRPV4 (WT) (RDB20677) pFN21A HaloTag-TRPV4 (R616Q) (RDB20678) pCMV-SNAP-D4H (RDB20679) pCMV-EGFP-Cav-1 (RDB20680) |