Transient receptor potential (TRP) proteins form plasma-membrane cation channels that act as sensors for diverse cellular stimuli. Here, we report a novel activation mechanism mediated by cysteine S-nitrosylation in TRP channels. Recombinant TRPC1, TRPC4, TRPC5, TRPV1, TRPV3 and TRPV4 of the TRPC and TRPV families, which are commonly classified as receptor-activated channels and thermosensor channels, induce entry of Ca2+ into cells in response to nitric oxide (NO). Labeling and functional assays using cysteine mutants, together with membrane sidedness in activating reactive disulfides, show that cytoplasmically accessible Cys553 and nearby Cys558 are nitrosylation sites mediating NO sensitivity in TRPC5. The responsive TRP proteins have conserved cysteines on the same N-terminal side of the pore region. Notably, nitrosylation of native TRPC5 upon G protein–coupled ATP receptor stimulation elicits entry of Ca2+ into endothelial cells. These findings reveal the structural motif for the NO-sensitive activation gate in TRP channels and indicate that NO sensors are a new functional category of cellular receptors extending over different TRP families.
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We thank D.E. Clapham and C. Strubing for TRPC5-DN, T. Furukawa and M. Nishida for helpful discussions, E. Mori and M. Sasaki for expert experiments and T. Kurosaki for IP3 receptor–deficient DT40 cells. This study was supported by research grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan, the Japan Society for the Promotion of Science, and from the Mitsubishi Foundation.
The authors declare no competing financial interests.
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Yoshida, T., Inoue, R., Morii, T. et al. Nitric oxide activates TRP channels by cysteine S-nitrosylation. Nat Chem Biol 2, 596–607 (2006) doi:10.1038/nchembio821
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