Molecular mechanism for H(2)S-induced activation of K(ATP) channels.
Hydrogen sulfide (H(2)S) is an endogenous opener of K(ATP) channels in many different types of cells. However, the molecular mechanism for an interaction between H(2)S and K(ATP) channel proteins remains unclear. The whole-cell patch-clamp technique and mutagenesis approach were used to examine the effects of H(2)S on different K(ATP) channel subunits, rvKir6.1 and rvSUR1, heterologously expressed in HEK-293 cells. H(2)S stimulated coexpressed rvKir6.1/rvSUR1 K(ATP) channels, but had no effect on K(ATP) currents generated by rvKir6.1 expression alone. Intracellularly applied sulfhydryl alkylating agent (N-ethylmaleimide, NEM), oxidizing agent (chloramine T, CLT), and a disulfide bond-oxidizing enzyme (protein disulfide isomerase) did not alter H(2)S effects on this recombinant channels. CLT, but not NEM, inhibited basal rvKir6.1/rvSUR1 currents, and both abolished the stimulatory effects of H(2)S on K(ATP) currents, when applied extracellularly. After selective cysteine residues (C6S and C26S but not C1051S and C1057S) in the extracellular loop of rvSUR1 subunits were point-mutated, H(2)S lost its stimulatory effects on rvKir6.1/rvSUR1 currents. Our results demonstrate that H(2)S interacts with Cys6 and Cys26 residues of the extracellular N terminal of rvSUR1 subunit of K(ATP) channel complex. Direct chemical modification of rvSUR1 subunit protein constitutes a molecular mechanism for the activation of K(ATP) channels by H(2)S.
Jiang
Bo
B
Department of Physiology & Pathophysiology, College of Medicine, Fudan University, Shanghai, PR China.
Tang
Guanghua
G
Cao
Kun
K
Wu
Lingyun
L
Wang
Rui
R
D000393:Air Pollutants / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D002700:Chloramines / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D005033:Ethylmaleimide / Q000378:metabolism; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism / Q000494:pharmacology; D015640:Ion Channel Gating / Q000187:drug effects*; D054086:KATP Channels / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D016877:Oxidants / Q000378:metabolism; D018408:Patch-Clamp Techniques; D017354:Point Mutation; D011487:Protein Conformation; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism; D013439:Sulfhydryl Reagents / Q000378:metabolism; D014105:Tosyl Compounds / Q000378:metabolism
D000393:Air Pollutants; D002700:Chloramines; D054086:KATP Channels; D016877:Oxidants; D021122:Protein Subunits; D013439:Sulfhydryl Reagents; D014105:Tosyl Compounds; C016300:chloramine-T; D019704:Protein Disulfide-Isomerases; D003545:Cysteine; D005033:Ethylmaleimide; D006862:Hydrogen Sulfide