title: Hydrogen sulfide inhibits human BK(Ca) channels.
abstract: Hydrogen sulfide (H(2)S) is produced endogenously in many types of mammalian cells. Evidence is now accumulating to suggest that H(2)S is an endogenous signalling molecule, with a variety of molecular targets, including ion channels. Here, we describe the effects of H(2)S on the large conductance, calcium-sensitive potassium channel (BK(Ca)). This channel contributes to carotid body glomus cell excitability and oxygen-sensitivity. The experiments were performed on HEK 293 cells, stably expressing the human BK(Ca) channel alpha subunit, using patch-clamp in the inside-out configuration. The H(2)S donor, NaSH (100microM-10 mM), inhibited BK(Ca) channels in a concentration-dependent manner with an IC(50) of ca. 670microM. In contrast to the known effects of CO donors, the H(2)S donor maximally decreased the open state probability by over 50% and shifted the half activation voltage by more than +16mV. In addition, although 1 mM KCN completely suppressed CO-evoked channel activation, it was without effect on the H(2)S-induced channel inhibition, suggesting that the effects of CO and H(2)S were non-competitive. RT-PCR showed that mRNA for both of the H(2)S-producing enzymes, cystathionine-beta-synthase and cystathionine-gamma-lyase, were expressed in HEK 293 cells and in rat carotid body. Furthermore, immunohistochemistry was able to localise cystathionine-gamma-lyase to glomus cells, indicating that the carotid body has the endogenous capacity to produce H(2)S. In conclusion, we have shown that H(2)S and CO have opposing effects on BK(Ca)channels, suggesting that these gases have separate modes of action and that they modulate carotid body activity by binding at different motifs in the BK(Ca)alphasubunit.
authors.affiliation: School of Bioscience, Museum Avenue, Cardiff University, Cardiff CF10 3AX, UK. telezhkinv@Cardiff.ac.uk
authors.forename: V
authors.initials: V
authors.lastname: Telezhkin
authors.forename: S P
authors.initials: SP
authors.lastname: Brazier
authors.forename: S
authors.initials: S
authors.lastname: Cayzac
authors.forename: C T
authors.initials: CT
authors.lastname: Müller
authors.forename: D
authors.initials: D
authors.lastname: Riccardi
authors.forename: P J
authors.initials: PJ
authors.lastname: Kemp
mesh_terms: D000818:Animals; D055592:Biophysical Phenomena / Q000187:drug effects; D002248:Carbon Monoxide / Q000494:pharmacology; D002344:Carotid Body / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D002460:Cell Line; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism / Q000494:pharmacology*; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / Q000037:antagonists & inhibitors*; D026902:Potassium Channel Blockers / Q000378:metabolism / Q000494:pharmacology*; D051381:Rats; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors
chemical_list: D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D026902:Potassium Channel Blockers; D011994:Recombinant Proteins; D002248:Carbon Monoxide; D006862:Hydrogen Sulfide