title: Mechanism for modulation of gating of connexin26-containing channels by taurine.
abstract: The mechanisms of action of endogenous modulatory ligands of connexin channels are largely unknown. Previous work showed that protonated aminosulfonates (AS), notably taurine, directly and reversibly inhibit homomeric and heteromeric channels that contain Cx26, a widely distributed connexin, but not homomeric Cx32 channels. The present study investigated the molecular mechanisms of connexin channel modulation by taurine, using hemichannels and junctional channels composed of Cx26 (homomeric) and Cx26/Cx32 (heteromeric). The addition of a 28-amino acid "tag" to the carboxyl-terminal domain (CT) of Cx26 (Cx26(T)) eliminated taurine sensitivity of homomeric and heteromeric hemichannels in cells and liposomes. Cleavage of all but four residues of the tag (Cx26(Tc)) resulted in taurine-induced pore narrowing in homomeric hemichannels, and restored taurine inhibition of heteromeric hemichannels (Cx26(Tc)/Cx32). Taurine actions on junctional channels were fully consistent with those on hemichannels. Taurine-induced inhibition of Cx26/Cx32(T) and nontagged Cx26 junctional channels was blocked by extracellular HEPES, a blocker of the taurine transporter, confirming that the taurine-sensitive site of Cx26 is cytoplasmic. Nuclear magnetic resonance of peptides corresponding to Cx26 cytoplasmic domains showed that taurine binds to the cytoplasmic loop (CL) and not the CT, and that the CT and CL directly interact. ELISA showed that taurine disrupts a pH-dependent interaction between the CT and the CT-proximal half of the CL. These studies reveal that AS disrupt a pH-driven cytoplasmic interdomain interaction in Cx26-containing channels, causing closure, and that the Cx26CT has a modulatory role in Cx26 function.
authors.affiliation: Department of Pharmacology and Physiology, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, USA. lockeda@gmail.com
authors.forename: Darren
authors.initials: D
authors.lastname: Locke
authors.forename: Fabien
authors.initials: F
authors.lastname: Kieken
authors.forename: Liang
authors.initials: L
authors.lastname: Tao
authors.forename: Paul L
authors.initials: PL
authors.lastname: Sorgen
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Harris
mesh_terms: D000072259:Connexin 26; D017630:Connexins / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D006531:HEPES / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007365:Intercellular Junctions / Q000378:metabolism; D055503:Protein Multimerization; D013654:Taurine / Q000494:pharmacology*; D000097002:Gap Junction beta-1 Protein
chemical_list: D000072259:Connexin 26; D017630:Connexins; D006531:HEPES; D013654:Taurine; D000097002:Gap Junction beta-1 Protein; C000607042:GJB2 protein, human