Gating of cyclic nucleotide-gated (CNGA1) channels by cGMP jumps and depolarizing voltage steps.
We expressed rod-type homotetrameric cyclic nucleotide-gated (CNGA1) channels in Xenopus oocytes and studied activation by photolysis-induced jumps of the 3',5'-cyclic guanosine monophosphate (cGMP) concentration and by voltage steps. cGMP jumps to increasing concentrations up to the EC50 value of 46.5 microM decelerate the activation gating, indicative that even at concentrations of cGMP << EC50 binding is not rate limiting. Above the EC50 value, activation by cGMP jumps is again accelerated to the higher concentrations. At the same cGMP concentration, the speed of the activation gating by depolarizing voltage steps is roughly similar to that by cGMP jumps. Permeating ions passing the pore more slowly (Rb+ > K+ > Na+) slow down the activation time course. At the single-channel level, cGMP jumps to high concentrations cause openings directly to the main open level without passing sublevels. From these results it is concluded that at both low and high cGMP the gating of homotetrameric CNGA1 channels is not rate-limited by the cGMP binding but by conformational changes of the channel which are voltage dependent and include movements in the pore region.
Institut für Physiologie II, Friedrich-Schiller-Universität Jena, Jena, Germany.
Vasilica
V
Nache
Jana
J
Kusch
Volker
V
Hagen
Klaus
K
Benndorf
D000494:Allosteric Regulation; D000818:Animals; D002417:Cattle; D006152:Cyclic GMP / Q000378:metabolism / Q000502:physiology*; D054815:Cyclic Nucleotide-Gated Cation Channels; D005260:Female; D015640:Ion Channel Gating / Q000502:physiology*; D007473:Ion Channels / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D009865:Oocytes / Q000378:metabolism / Q000502:physiology; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D012413:Rubidium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D013997:Time Factors; D014982:Xenopus laevis
D054815:Cyclic Nucleotide-Gated Cation Channels; D007473:Ion Channels; D012964:Sodium; D006152:Cyclic GMP; D012413:Rubidium; D011188:Potassium