TRPC1 forms the stretch-activated cation channel in vertebrate cells.
The mechanosensitive cation channel (MscCa) transduces membrane stretch into cation (Na(+), K(+), Ca(2+) and Mg(2+)) 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 (>or=10(7)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 (M(r) 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.
Maroto
Rosario
R
Department of Neuroscience & Cell Biology, UTMB, Galveston, TX 77555, USA.
Raso
Albert
A
Wood
Thomas G
TG
Kurosky
Alex
A
Martinac
Boris
B
Hamill
Owen P
OP
D000818:Animals; D016466:CHO Cells; D015220:Calcium Channels / Q000502:physiology*; D006224:Cricetinae; D006801:Humans; D007473:Ion Channels / Q000737:chemistry*; D008081:Liposomes; D051379:Mice; D018408:Patch-Clamp Techniques; D016372:RNA, Antisense / Q000494:pharmacology; D013314:Stress, Mechanical; D000074025:TRPA1 Cation Channel; D050052:TRPC Cation Channels; D014162:Transfection; D050051:Transient Receptor Potential Channels; D014982:Xenopus laevis
D015220:Calcium Channels; D007473:Ion Channels; D008081:Liposomes; D016372:RNA, Antisense; D000074025:TRPA1 Cation Channel; D050052:TRPC Cation Channels; D050051:Transient Receptor Potential Channels; C475685:Trpa1 protein, mouse; C096351:transient receptor potential cation channel, subfamily C, member 1