Diversity of amyloid β protein fragment [1-40]-formed channels

1. The lipid bilayer technique was used to characterize the biophysical and pharmacological properties of several ion channels formed by incorporating amyloid beta protein fragment (A betaP) 1-40 into lipid membranes. Based on the conductance, kinetics, selectivity, and pharmacological properties, the following A betaP [1-40]-formed ion channels have been identified: (i) The A betaP[1-40]-formed bursting fast cation channel was characterized by (a) a single channel conductance of 63 pS (250/50 mM KCl cis/trans) at +140 mV, 17 pS (250/50 mM KC/ cis/trans) at -160 mV, and the nonlinear current-voltage relationship drawn to a third-order polynomial, (b) selectivity sequence P-K>P-Na>P-Li = 1.0:0.60:0.47, (c) P-o of 0.22 at 0 mV and 0.55 at +120 mV, and (d) Zn2+-induced reduction in current amplitude, a typical property of a slow block mechanism. (ii) The A betaP[1-40]-formed spiky fast cation channel was characterized by (a) a similar kinetics to the bursting fast channel with exception for the absence of the long intraburst closures, (b) single channel conductance of 63 pS (250/50 KCI) at +140 mV 17 pS (250/50 KCI) at -160 mV, the current-voltage relationship nonlinear drawn to a third-order polynomial fit, and (c) selectivity sequence P-Rb>P-K>P-Cs>P-Na>P-Li = 1.3:1.0:0.46:0.40:027. (iii) The A betaP[1-40]-formed medium conductance channel was characterized by (a) 275 pS (250/50 mM KCL cis/trans) at +140 mV and 19 pS (250/50 mM KCl cis/trans) at -160 mV and (b) inactivation at V(m)s more negative than -120 and more positive than + 120 mV. (iv) The A betaP[1-40] -formed inactivating large conductance channel was characterized by (a) fast and slow modes of opening to seven multilevel conductances ranging between 0-589 pS (in 250/50 mM KCl) at +140 mV and 0-704 pS (in 250/50 mM KCI) at -160 mV, (b)The fast mode which had a conductance of <250 pS was voltage dependent. The inactivation was described by a bell-shaped curve with a peak lag time of 7.2 s at +36 mV. The slow mode which had a conductance of >250 pS was also voltage dependent. The inactivation was described by a bell-shaped curve with a peak lag time of 7.0 s at -76 mV, (c) the value of P-K/P-choline for the fast mode was 3.9 and selectivity sequence P-K>F-Cs>P-Na>P-Li = 1.0:0.94:0.87:0.59. The value of P-K/P-choline for the blow mode was 2.7 and selectivity sequence P-K>P-Na>P-Li>P-Cs = 1.0:0.59:0.49:0.21, and (d) asymmetric blockade with 10 mM Zn2+-induced reduction in the large conductance state of the slow mode mediated via slow block mechanism. The fast mode of the large conductance channel was not affected by 10 mM Zn2+. 2. It has been suggested that, although the bursting fast channel, the spiky fast channel and the inactivating medium conductance channel are distinct, it is possible that they are intermediate configurations of yet another configuration underlying the inactivating large conductance channel. It is proposed that this heterogeneity is one of the most common features of these positively-charged cytotoxic amyloid-formed channels reflecting these channels ability to modify multiple cellular functions. 3. Furthermore, the formation of beta -sheet based oligomers could be an important common step in the formation of cytotoxic amyloid channels.