title: Functional interactions between A' helices in the C-linker of open CNG channels.
abstract: Cyclic nucleotide-gated (CNG) channels are nonselective cation channels that are activated by the direct binding of the cyclic nucleotides cAMP and cGMP. The region linking the last membrane-spanning region (S6) to the cyclic nucleotide binding domain in the COOH terminus, termed the C-linker, has been shown to play an important role in coupling cyclic nucleotide binding to opening of the pore. In this study, we explored the intersubunit proximity between the A' helices of the C-linker regions of CNGA1 in functional channels using site-specific cysteine substitution. We found that intersubunit disulfide bonds can be formed between the A' helices in open channels, and that inducing disulfide bonds in most of the studied constructs resulted in potentiation of channel activation. This suggests that the A' helices of the C-linker regions are in close proximity when the channel is in the open state. Our finding is not compatible with a homology model of the CNGA1 C-linker made from the recently published X-ray crystallographic structure of the hyperpolarization-activated, cyclic nucleotide-modulated (HCN) channel COOH terminus, and leads us to suggest that the C-linker region depicted in the crystal structure may represent the structure of the closed state. The opening conformational change would then involve a movement of the A' helices from a position parallel to the axis of the membrane to one perpendicular to the axis of the membrane.
authors.affiliation: Department of Phisiology and Biophysics, University of Washington, Seattle, WA 98195, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Hua
authors.forename: Sharona E
authors.initials: SE
authors.lastname: Gordon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D054815:Cyclic Nucleotide-Gated Cation Channels; D004220:Disulfides / Q000737:chemistry; D004229:Dithiothreitol / Q000494:pharmacology; D005260:Female; D007473:Ion Channels / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes; D010618:Phenanthrolines / Q000037:antagonists & inhibitors / Q000494:pharmacology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D014981:Xenopus
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; D004220:Disulfides; D007473:Ion Channels; D010618:Phenanthrolines; D004229:Dithiothreitol; C025205:1,10-phenanthroline
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