Disruption of membrane cholesterol organization impairs the activity of PIEZO1 channel clusters
| Autor: | Pietro Ridone, Boris Martinac, Massimo Vassalli, Alexander Macmillan, Philip A. Gottlieb, Elvis Pandzic, Charles D. Cox |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Physiology Cholesterol PIEZO1 Biophysics Lipids and Membranes Fusion protein Article Cell membrane 03 medical and health sciences chemistry.chemical_compound Electrophysiology 030104 developmental biology 0302 clinical medicine Membrane Mechanosensitive ion channel medicine.anatomical_structure chemistry medicine Cellular Physiology 030217 neurology & neurosurgery Homeostasis |
| Zdroj: | The Journal of General Physiology |
| ISSN: | 0022-1295 |
| Popis: | The essential mammalian mechanosensitive channel PIEZO1 organizes in the plasma membrane into nanometric clusters that depend on the integrity of cholesterol domains to rapidly detect applied force and especially inactivate synchronously, the most commonly altered feature of PIEZO1 in pathology. The human mechanosensitive ion channel PIEZO1 is gated by membrane tension and regulates essential biological processes such as vascular development and erythrocyte volume homeostasis. Currently, little is known about PIEZO1 plasma membrane localization and organization. Using a PIEZO1-GFP fusion protein, we investigated whether cholesterol enrichment or depletion by methyl-β-cyclodextrin (MBCD) and disruption of membrane cholesterol organization by dynasore affects PIEZO1-GFP’s response to mechanical force. Electrophysiological recordings in the cell-attached configuration revealed that MBCD caused a rightward shift in the PIEZO1-GFP pressure–response curve, increased channel latency in response to mechanical stimuli, and markedly slowed channel inactivation. The same effects were seen in native PIEZO1 in N2A cells. STORM superresolution imaging revealed that, at the nanoscale, PIEZO1-GFP channels in the membrane associate as clusters sensitive to membrane manipulation. Both cluster distribution and diffusion rates were affected by treatment with MBCD (5 mM). Supplementation of polyunsaturated fatty acids appeared to sensitize the PIEZO1-GFP response to applied pressure. Together, our results indicate that PIEZO1 function is directly dependent on the membrane composition and lateral organization of membrane cholesterol domains, which coordinate the activity of clustered PIEZO1 channels. |
| Databáze: | OpenAIRE |
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