TRPC1 forms the stretch-activated cation channel in vertebrate cells

Rosario Maroto, Albert Raso, Thomas Wood, Alex Kurosky, Boris Martinac, Owen Hamill

Research output: Contribution to journalArticle

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Abstract

The mechanosensitive cation channel (MscCa) transduces membrane stretch into cation (Na+, K+, Ca2+ and Mg2+) flux across the cell membrane, and is implicated in cell-volume regulation, cell locomotion, muscle dystrophy and cardiac arrhythmias. However, the membrane protein(s) that form the MscCa in vertebrates remain unknown. Here, we use an identification strategy that is based on detergent solubilization of frog oocyte membrane proteins, followed by liposome reconstitution and evaluation by patch-clamp. The oocyte was chosen because it expresses the prototypical MscCa (≥107 MscCa/oocyte) that is preserved in cytoskeleton-deficient membrane vesicles. We identified a membrane-protein fraction that reconstituted high MscCa activity and showed an abundance of a protein that had a relative molecular mass of 80,000 (Mr 80K). This protein was identified, by immunological techniques, as the canonical transient receptor potential channel 1 (TRPC1). Heterologous expression of the human TRPC1 resulted in a >1,000% increase in MscCa patch density, whereas injection of a TRPC1-specific antisense RNA abolished endogenous MscCa activity. Transfection of human TRPC1 into CHO-K1 cells also significantly increased MscCa expression. These observations indicate that TRPC1 is a component of the vertebrate MscCa, which is gated by tension developed in the lipid bilayer, as is the case in various prokaryotic mechanosensitive (MS) channels.

Original languageEnglish (US)
Pages (from-to)179-185
Number of pages7
JournalNature Cell Biology
Volume7
Issue number2
DOIs
StatePublished - Feb 2005

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Transient Receptor Potential Channels
Vertebrates
Cations
Oocytes
Membrane Proteins
Immunologic Techniques
Antisense RNA
CHO Cells
Lipid Bilayers
Cytoskeleton
Ion Channels
Cell Size
Liposomes
Anura
Detergents
Cell Movement
Transfection
Cardiac Arrhythmias
Proteins
Cell Membrane

ASJC Scopus subject areas

  • Cell Biology

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TRPC1 forms the stretch-activated cation channel in vertebrate cells. / Maroto, Rosario; Raso, Albert; Wood, Thomas; Kurosky, Alex; Martinac, Boris; Hamill, Owen.

In: Nature Cell Biology, Vol. 7, No. 2, 02.2005, p. 179-185.

Research output: Contribution to journalArticle

Maroto, R, Raso, A, Wood, T, Kurosky, A, Martinac, B & Hamill, O 2005, 'TRPC1 forms the stretch-activated cation channel in vertebrate cells', Nature Cell Biology, vol. 7, no. 2, pp. 179-185. https://doi.org/10.1038/ncb1218
Maroto, Rosario ; Raso, Albert ; Wood, Thomas ; Kurosky, Alex ; Martinac, Boris ; Hamill, Owen. / TRPC1 forms the stretch-activated cation channel in vertebrate cells. In: Nature Cell Biology. 2005 ; Vol. 7, No. 2. pp. 179-185.
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