Cyclic nucleotide-gated channels of rat olfactory receptor cells: Divalent cations control the sensitivity to cAMP

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Abstract

cAMP-gated channels were studied in inside-out membrane patches excised from the apical cellular pole of isolated olfactory receptor cells of the rat. In the absence of divalent cations the dose-response curve of activation of patch current by cAMP had a K(M) of 4.0 μM at -50 mV and of 2.5 μM at +50 mV. However, addition of 0.2 or 0.5 mM Ca2+ shifted the K(M) of cAMP reversibly to the higher cAMP concentrations of 33 or 90 μM, respectively, at -50 mV. Among divalent cations, the relative potency for inducing cAMP affinity shifts was: Ca2+ > Sr2+ > Mn2+ > Ba2+ > Mg2+, of which Mg2+ (up to 3 mM) did not shift the K(M) at all. This potency sequence corresponds closely to that required for the activation of calmodulin. However, the Ca2+-sensitivity is lower than expected for a calmodulin- mediated action. Brief (60 s) transient exposure to 3 mM Mg2+, in the absence of other divalent cations, had a protective effect in that following washout of Mg2+, subsequent exposure to 0.2 mM Ca2+ no longer caused affinity shifts. This protection effect did not occur in intact cells and was probably a consequence of patch excision, possibly representing ablation of a regulatory protein from the channel cyclic nucleotide binding site. Thus, the binding of divalent cations, probably via a regulatory protein, controls the sensitivity of the cAMP-gated channels to cAMP. The influx of Ca2+ through these channels during the odorant response may rise to a sufficiently high concentration at the intracellular membrane surface to contribute to the desensitization of the odorant-induced response. The results also indicate that divalent cation effects on cyclic nucleotide-gated channels may depend on the sequence of pre-exposure to other divalent cations.

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Lynch, J. W., & Lindemann, B. (1994). Cyclic nucleotide-gated channels of rat olfactory receptor cells: Divalent cations control the sensitivity to cAMP. Journal of General Physiology, 103(1), 87–106. https://doi.org/10.1085/jgp.103.1.87

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