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of outstanding interest. The authors used biochemical analysis, molecular cloning and functional expression to identify a 240 kDa protein as the complete β subunit of the rod CNG channel. This subunit is shown to consist of a β′ part, which is homologous to subunit 2 of the human rod channel [9], and a glutamic-acid-rich domain related to a previously cloned protein of unknown function [24]. The β′ part contains the binding site for calmodulin and when co-expressed with the α subunit forms a hetero-oligomeric channel with molecular and electrophysiological properties similar to the native rod CNG channel (see also [9,22]). The function of the glutamic-acid-rich part is currently unknown.
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2+ coordinates between neighboring subunits of the channel to stabilize the open state of the channel. Functional expression of tandem dimers also supports the view that the homomultimeric rod CNG channel probably exists as a tetramer.
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Molecular mechanism for ligand discrimination of cyclic nucleotide-gated channels
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of outstanding interest. Using site-directed mutagenesis, the authors identify aspartic acid residue Asp604 within the cyclic-nucleotide-binding site of the rod channel as the main determinant responsible for the high degree of selectivity of the rod channel for cGMP. They proposes a model based on the known X-ray crystallographic structure of the CAP protein, in which aspartic acid Asp604 forms a pair of hydrogen bonds with the nitrogens of the guanine ring of cGMP, thereby facilitating the allosteric transition of the channel to its open state.
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Varnum MD, Black KD, Zagotta WN. Molecular mechanism for ligand discrimination of cyclic nucleotide-gated channels. of outstanding interest Neuron. 15:1995;619-625 Using site-directed mutagenesis, the authors identify aspartic acid residue Asp604 within the cyclic-nucleotide-binding site of the rod channel as the main determinant responsible for the high degree of selectivity of the rod channel for cGMP. They proposes a model based on the known X-ray crystallographic structure of the CAP protein, in which aspartic acid Asp604 forms a pair of hydrogen bonds with the nitrogens of the guanine ring of cGMP, thereby facilitating the allosteric transition of the channel to its open state.
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Varnum, M.D.1
Black, K.D.2
Zagotta, W.N.3
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Covalent activation of retinal rod cGMP-gated channels reveals a functional heterogeneity in the ligand binding sites
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of outstanding interest. The cGMP photoaffinity analogue 8-p-azidophenacylthio-cGMP is used to irreversibly label all but one cGMP-binding site on the rod CNG channel in excised patches. Dose-response curves reveal the presence of two populations of sites differing in their apparent affinity for cGMP by a factor of ~ 25. This heterogeneity appears to result at least in part from sequence differences in the cGMP-binding site for the α and β subunits. This method should prove to be valuable for future studies of channel modulation by calmodulin or other mechanism at the level of individual cGMP-binding sites.
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Karpen JW, Brown RL. Covalent activation of retinal rod cGMP-gated channels reveals a functional heterogeneity in the ligand binding sites. of outstanding interest J Gen Physiol. 107:1996;169-181 The cGMP photoaffinity analogue 8-p-azidophenacylthio-cGMP is used to irreversibly label all but one cGMP-binding site on the rod CNG channel in excised patches. Dose-response curves reveal the presence of two populations of sites differing in their apparent affinity for cGMP by a factor of ~ 25. This heterogeneity appears to result at least in part from sequence differences in the cGMP-binding site for the α and β subunits. This method should prove to be valuable for future studies of channel modulation by calmodulin or other mechanism at the level of individual cGMP-binding sites.
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Localization of regions affecting an allosteric transition in cyclic nucleotide-activated channels
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2+ to this residue results in inhibition of cyclic nucleotide activation of the olfactory channel by stabilizing the closed state of the channel.
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2+ to this residue results in inhibition of cyclic nucleotide activation of the olfactory channel by stabilizing the closed state of the channel.
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Gordon, S.E.1
Zagotta, W.N.2
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A histidine residue associated with the gate of the cyclic nucleotide-activated channels in rod photoreceptors
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2+ to this residue results in potentiation of cyclic nucleotide activation of the rod channel by stabilizing the open state.
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2+ to this residue results in potentiation of cyclic nucleotide activation of the rod channel by stabilizing the open state.
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Gordon, S.E.1
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of outstanding interest. The accessibility of cysteine residues substituted at various sites within the pore region of the rod CNG channel has been investigated by applying charged sulfhydryl reagents to the inside and outside of membrane patches. On the basis of these results, the authors have suggested that the pore region of each subunit consists of a loop that extends toward the central axis of the channel and, together with the pore regions of the other subunits, forms the ion selectivity filter and gate of the channel. Structural changes in the channel gate occur as the channel is converted from its closed state to its open state.
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Sun Z-P, Akabas MH, Goulding EH, Karlin A, Siegelbaum SA. Exposure of residues in the cyclic nucleotide-gated channel pore: P region structure and function in gating. of outstanding interest Neuron. 16:1996;141-149 The accessibility of cysteine residues substituted at various sites within the pore region of the rod CNG channel has been investigated by applying charged sulfhydryl reagents to the inside and outside of membrane patches. On the basis of these results, the authors have suggested that the pore region of each subunit consists of a loop that extends toward the central axis of the channel and, together with the pore regions of the other subunits, forms the ion selectivity filter and gate of the channel. Structural changes in the channel gate occur as the channel is converted from its closed state to its open state.
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Sun Z-P1
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0029117775
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2+-mediated excitation and adaptation mechanisms of rods, cones, and olfactory neurons
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2+-mediated excitation and adaptation mechanisms of rods, cones, and olfactory neurons.
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2+ pump and myosin light-chain kinase is described. This generalized mechanism of regulation is related to the mechanism by which calmodulin modulates the apparent affinity of the olfactory for cAMP see ([43]).
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2+ pump and myosin light-chain kinase is described. This generalized mechanism of regulation is related to the mechanism by which calmodulin modulates the apparent affinity of the olfactory for cAMP see ([43]).
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