Calcium dependence of C-type natriuretic peptide-formed fast K(+) channel.

Abstract

The lipid bilayer technique was used to characterize the Ca(2+) dependence of a fast K(+) channel formed by a synthetic 17-amino acid segment [OaCNP-39-(1-17)] of a 39-amino acid C-type natriuretic peptide (OaCNP-39) found in platypus (Ornithorhynchus anatinus) venom (OaV). The OaCNP-39-(1-17)-formed K(+) channel was reversibly dependent on 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-buffered cis (cytoplasmic) Ca(2+) concentration ([Ca(2+)](cis)). The channel was fully active when [Ca(2+)](cis) was >10(-4) M and trans (luminal) Ca(2+) concentration was 1.0 mM, but not at low [Ca(2+)](cis). The open probability of single channels increased from zero at 1 x 10(-6) M cis Ca(2+) to 0.73 +/- 0.17 (n = 22) at 10(-3) M cis Ca(2+). Channel openings to the maximum conductance of 38 pS were rapidly and reversibly activated when [Ca(2+)](cis), but not trans Ca(2+) concentration (n = 5), was increased to >5 x 10(-4) M (n = 14). Channel openings to the submaximal conductance of 10.5 pS were dominant at >/=5 x 10(-4) M Ca(2+). K(+) channels did not open when cis Mg(2+) or Sr(2+) concentrations were increased from zero to 10(-3) M or when [Ca(2+)](cis) was maintained at 10(-6) M (n = 3 and 2). The Hill coefficient and the inhibition constant were 1 and 0.8 x 10(-4) M cis Ca(2+), respectively. This dependence of the channel on high [Ca(2+)](cis) suggests that it may become active under 1) physiological conditions where Ca(2+) levels are high, e.g., during cardiac and skeletal muscle contractions, and 2) pathological conditions that lead to a Ca(2+) overload, e.g., ischemic heart and muscle fatigue. The channel could modify a cascade of physiological functions that are dependent on the Ca(2+)-activated K(+) channels, e.g., vasodilation and salt secretion.