A component of platypus (Ornithorhynchus anatinus) venom forms slow-kinetic cation channels.
The lipid bilayer technique is used to examine the biophysical properties of anion and cation channels frequently formed by platypus (Ornithorhynchus anatinus) venom (OaV). The OaV-formed anion channel in 250/50 mm KCl cis/trans has a maximum conductance of 857 +/- 23 pS (n = 5) in 250/50 mm KCl cis/trans. The current-voltage relationship of this channel shows strong inward rectification. The channel activity undergoes time-dependent inactivation that can be removed by depolarizing voltage steps more positive than the reversal potential for chloride, E(Cl), (+40 mV). The reversal potential of the OaV-formed slow current activity in 250/50 mm KCl cis/trans is close to the potassium equilibrium potential (E(K)) of -40 mV. The conductance values for the slow channel are 22.5 +/- 2.6 pS and 41.38 +/- 4.2 pS in 250/50 and 750/50 mm cis/trans, respectively. The gating kinetics of the slow ion channels are voltage-dependent. The channel open probability (P(o)) is between 0.1 and 0.8 at potentials between 0 and +140 mV. The channel frequency (F(o)) increases with depolarizing voltages between 0 and +140 mV, whereas mean open time (T(o)) and mean closed time (T(c)) decrease. Ion substitution experiments of the cis solution show that the channel has conductance values of 21.47 +/- 2. 3 and 0.53 +/- 0.1 pS in 250 mm KCl and choline Cl, respectively. The amplitude of the single channel current is dependent on [K(+)](cis) and the current reversal potential (E(rev)) responds to increases in [K(+)](cis) by shifting to more negative voltages. The increase in current amplitude as a function of increasing [K(+)](cis) can be best described by a third order polynomial fit. At +140 mV, the values of the maximal single channel conductance (gamma(max)) and the concentration for half maximal gamma (K(s)) are 38.6 pS and 380 mm and decline to 15.76 pS and 250 mm at 0 mV, respectively. The ion selectivity of the channel to K(+), Na(+), Cs(+) and choline(+) was determined in ion substitution experiments. The permeability values for P(K(+)):P(Na(+)):P(Cs(+)):P(choline(+)) were 1:1:0.63:0.089, respectively. On the other hand, the activity of the slow channel was eliminated (Fig. 7B). The slow channel was reversibly inhibited by [TEA(+)](trans) and the half-maximal inhibitory concentration (K(i)) was approximately 48 mm.
Kourie
J I
JI
Membrane Transport Group, Department of Chemistry, The Faculties, The Australian National University, Canberra City, ACT, 0200 Australia.
D000818:Animals; D066298:In Vitro Techniques; D007473:Ion Channels / Q000187:drug effects* / Q000378:metabolism; D007700:Kinetics; D008051:Lipid Bilayers; D008564:Membrane Potentials / Q000187:drug effects; D020098:Natriuretic Peptide, C-Type / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry / Q000633:toxicity; D010987:Platypus; D014688:Venoms / Q000737:chemistry / Q000633:toxicity*
D007473:Ion Channels; D008051:Lipid Bilayers; D010455:Peptides; D014688:Venoms; D020098:Natriuretic Peptide, C-Type