Effects of ultraviolet modification on the gating energetics of cyclic nucleotide-gated channels.
Middendorf et al. (Middendorf, T.R., R.W. Aldrich, and D.A. Baylor. 2000. J. Gen. Physiol. 116:227-252) showed that ultraviolet light decreases the current through cloned cyclic nucleotide-gated channels from bovine retina activated by high concentrations of cGMP. Here we probe the mechanism of the current reduction. The channels' open probability before irradiation, P(o)(0), determined the sign of the change in current amplitude that occurred upon irradiation. UV always decreased the current through channels with high initial open probabilities [P(o)(0) > 0.3]. Manipulations that promoted channel opening antagonized the current reduction by UV. In contrast, UV always increased the current through channels with low initial open probabilities [P(o)(0) < or = 0.02], and the magnitude of the current increase varied inversely with P(o)(0). The dual effects of UV on channel currents and the correlation of both effects with P(o)(0) suggest that the channels contain two distinct classes of UV target residues whose photochemical modification exerts opposing effects on channel gating. We present a simple model based on this idea that accounts quantitatively for the UV effects on the currents and provides estimates for the photochemical quantum yields and free energy costs of modifying the UV targets. Simulations indicate that UV modification may be used to produce and quantify large changes in channel gating energetics in regimes where the associated changes in open probability are not measurable by existing techniques.
Neurobiology Department, Howard Hughes Medical Institute, School of Medicine, Stanford University, California 94305, USA.
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Aldrich
D000818:Animals; D002417:Cattle; D003198:Computer Simulation; D006152:Cyclic GMP / Q000494:pharmacology; D054815:Cyclic Nucleotide-Gated Cation Channels; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004734:Energy Metabolism / Q000528:radiation effects; D015640:Ion Channel Gating / Q000187:drug effects / Q000528:radiation effects*; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D008564:Membrane Potentials / Q000187:drug effects / Q000528:radiation effects; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D010775:Photic Stimulation; D010777:Photochemistry; D012160:Retina / Q000737:chemistry; D014364:Tryptophan / Q000737:chemistry; D014466:Ultraviolet Rays; D014981:Xenopus
D054815:Cyclic Nucleotide-Gated Cation Channels; D007473:Ion Channels; D008024:Ligands; D014364:Tryptophan; D006152:Cyclic GMP