Identification of apamin binding sites in rat intestinal mucosa

Apamin, a specific blocker of one class of Ca(2+)-activated K+ channes, was used to detect the apamin receptors associated with K+ channels in the mucosa of the rat jejunum and colon. Two receptor sites for 125I-apamin have been identified. These sites differed in their affinity for apamin (jejunum: KD1 = 1.1 nM and KD2 = 170 nM; colon: KD1 = 0.5 nM and KD2 = 1.1 nM and KD2 = 140 nM) and the maximum number of sites (jejunum: B(max1) = 111 and B(max2) = 4030; colon: B(max1) = 187 and B(max2) = 7550 fmol/mg of protein). 125I-apamin binding was stimulated by K+ ions with K0.5 = 1.0 mM and inhibited by the neuromuscular blocker tubocurarine (KI = 50 microM). We interpret these data to demonstrate that the high-affinity, low-capacity binding sites reflect the existence of apamin-sensitive K+ channels in the intestinal mucosa.