Gaston, BenjaminSmith, LauraBosch, JürgenSeckler, JamesKunze, DianaKiselar, JannaMarozkina, NadzeyaHodges, Craig A.Wintrobe, PatrickMcGee, KellenMorozkina, Tatiana S.Burton, Spencer T.Lewis, TristanStrassmaier, TimothyGetsy, PaulinaBates, James N.Lewis, Stephen J.2021-08-092021-08-092020-08-13Gaston, B., Smith, L., Bosch, J., Seckler, J., Kunze, D., Kiselar, J., Marozkina, N., Hodges, C. A., Wintrobe, P., McGee, K., Morozkina, T. S., Burton, S. T., Lewis, T., Strassmaier, T., Getsy, P., Bates, J. N., & Lewis, S. J. (2020). Voltage-gated potassium channel proteins and stereoselective S-nitroso-l-cysteine signaling. JCI Insight, 5(18). https://doi.org/10.1172/jci.insight.1341740021-9738https://hdl.handle.net/1805/26351S-nitroso-l-cysteine (L-CSNO) behaves as a ligand. Its soluble guanylate cyclase–independent (sGC-independent) effects are stereoselective — that is, not recapitulated by S-nitroso-d-cysteine (D-CSNO) — and are inhibited by chemical congeners. However, candidate L-CSNO receptors have not been identified. Here, we have used 2 complementary affinity chromatography assays — followed by unbiased proteomic analysis — to identify voltage-gated K+ channel (Kv) proteins as binding partners for L-CSNO. Stereoselective L-CSNO–Kv interaction was confirmed structurally and functionally using surface plasmon resonance spectroscopy; hydrogen deuterium exchange; and, in Kv1.1/Kv1.2/Kvβ2-overexpressing cells, patch clamp assays. Remarkably, these sGC-independent L-CSNO effects did not involve S-nitrosylation of Kv proteins. In isolated rat and mouse respiratory control (petrosyl) ganglia, L-CSNO stereoselectively inhibited Kv channel function. Genetic ablation of Kv1.1 prevented this effect. In intact animals, L-CSNO injection at the level of the carotid body dramatically and stereoselectively increased minute ventilation while having no effect on blood pressure; this effect was inhibited by the L-CSNO congener S-methyl-l-cysteine. Kv proteins are physiologically relevant targets of endogenous L-CSNO. This may be a signaling pathway of broad relevance.en-USAttribution 4.0 InternationalRespirationRespirationCysteineGangliaPotassium ChannelsVoltage-GatedS-NitrosothiolsArticle