title: All-trans-retinal is a closed-state inhibitor of rod cyclic nucleotide-gated ion channels.
abstract: Rod vision begins when 11-cis-retinal absorbs a photon and isomerizes to all-trans-retinal (ATR) within the photopigment, rhodopsin. Photoactivated rhodopsin triggers an enzyme cascade that lowers the concentration of cGMP, thereby closing cyclic nucleotide-gated (CNG) ion channels. After isomerization, ATR dissociates from rhodopsin, and after a bright light, this release is expected to produce a large surge of ATR near the CNG channels. Using excised patches from Xenopus oocytes, we recently showed that ATR shuts down cloned rod CNG channels, and that this inhibition occurs in the nanomolar range (aqueous concentration) at near-physiological concentrations of cGMP. Here we further characterize the ATR effect and present mechanistic information. ATR was found to decrease the apparent cGMP affinity, as well as the maximum current at saturating cGMP. When ATR was applied to outside-out patches, inhibition was much slower and less effective than when it was applied to inside-out patches, suggesting that ATR requires access to the intracellular surface of the channel or membrane. The apparent ATR affinity and maximal inhibition of heteromeric (CNGA1/CNGB1) channels was similar to that of homomeric (CNGA1) channels. Single-channel and multichannel data suggest that channel inhibition by ATR is reversible. Inhibition by ATR was not voltage dependent, and the form of its dose-response relation suggested multiple ATR molecules interacting per channel. Modeling of the data obtained with cAMP and cGMP suggests that ATR acts by interfering with the allosteric opening transition of the channel and that it prefers closed, unliganded channels. It remains to be determined whether ATR acts directly on the channel protein or instead alters channel-bilayer interactions.
authors.affiliation: Department of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, RI 02912, USA.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: McCabe
authors.forename: Diana M
authors.initials: DM
authors.lastname: Pelosi
authors.forename: Michelle
authors.initials: M
authors.lastname: Tetreault
authors.forename: Andrew
authors.initials: A
authors.lastname: Miri
authors.forename: Wang
authors.initials: W
authors.lastname: Nguitragool
authors.forename: Pranisa
authors.initials: P
authors.lastname: Kovithvathanaphong
authors.forename: Rahul
authors.initials: R
authors.lastname: Mahajan
authors.forename: Anita L
authors.initials: AL
authors.lastname: Zimmerman
mesh_terms: D000818:Animals; D002417:Cattle; D003198:Computer Simulation; D006152:Cyclic GMP / Q000378:metabolism*; D054815:Cyclic Nucleotide-Gated Cation Channels; D004305:Dose-Response Relationship, Drug; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D007473:Ion Channels / Q000037:antagonists & inhibitors* / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008954:Models, Biological; D009865:Oocytes / Q000502:physiology; D017948:Retinal Rod Photoreceptor Cells / Q000502:physiology*; D014801:Vitamin A / Q000494:pharmacology*; D014982:Xenopus laevis
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; D007473:Ion Channels; D014801:Vitamin A; D006152:Cyclic GMP