RT Journal Article SR Electronic T1 Trans-nitrosylation Directs TRPA1 Selectivity in N-Nitrosamine Activators JF Molecular Pharmacology JO Mol Pharmacol FD American Society for Pharmacology and Experimental Therapeutics SP mol.113.088864 DO 10.1124/mol.113.088864 A1 Daisuke Kozai A1 Yoji Kabasawa A1 Maximilian Ebert A1 Shigeki Kiyonaka A1 Fir man A1 Yuko Otani A1 Tomohiro Numata A1 Nobuaki Takahashi A1 Yasuo Mori A1 Tomohiko Ohwada YR 2013 UL http://molpharm.aspetjournals.org/content/early/2013/11/07/mol.113.088864.abstract AB S-Nitrosylation, the addition of a nitrosyl group to cysteine thiols, regulates various protein functions to mediate nitric oxide (NO) bioactivity. Recent studies have demonstrated that selectivity in protein S-nitrosylation signaling pathways is conferred through trans-nitrosylation, a transfer of the NO group, between proteins via interaction. We previously demonstrated that sensitivity to activation by synthetic NO-releasing agents via S-nitrosylation is a common feature of members of the transient receptor potential (TRP) family of Ca2+-permeable cation channels. However, strategies to confer subtype selectivity to nitrosylating agents targeted to TRP channels are yet to be developed. Here, we show selective activation of TRPA1 channels by novel NO donors derived from the 7-azabenzobicyclo[2.2.1]heptane (ABBH) N-nitrosamines, which exhibit trans-nitrosylation reactivity to thiols without releasing NO. The N-nitroso-2-exo,3-exo-ditrifluoromethyl-7-azabenzobicyclo[2.2.1]heptane (NNO-ABBH1) elicits S-nitrosylation of TRPA1 proteins, and dose-dependently induces robust Ca2+ influx via both recombinant and native TRPA1 channels, but not via other NO-activated TRP channels. TRPA1 activation by NNO-ABBH1 is suppressed by specific cysteine mutations but not by NO scavenging, suggesting that cysteine trans-nitrosylation underlies the activation of TRPA1 by NNO-ABBH1. This is supported by the correlation of N-NO bond reactivity and TRPA1-activating potency in a congeneric series of ABBH N-nitrosamines. Interestingly, non-electrophilic derivatives of ABBH also activate TRPA1 selectively but less potently compared to NNO-ABBH1. Thus, ABBH N-nitrosamines confer subtype selectivity on S-nitrosylation in TRP channels through synergetic effects of two chemical processes: cysteine trans-nitrosylation and molecular recognition of the non-electrophilic moiety.