amyloid-beta-peptides and zinc-chloride

The disruption of copper homeostasis and the oxidative stress induced by Cu-amyloids are crucial features of Alzheimer's disease pathology. The copper specific N

Topics: Aminoquinolines; Amyloid beta-Peptides; Ascorbic Acid; Carrier Proteins; Chelating Agents; Chlorides; Copper; Homeostasis; Humans; Molecular Structure; Oxidation-Reduction; Oxidative Stress; Peptide Fragments; Reactive Oxygen Species; Zinc Compounds

In this paper, a kind of gold nanoparticle (GNP)-based colorimetric assay has been developed for studying the reversible interaction of β-amyloid peptide (Aβ) with Cu(2+) and Zn(2+), and quantitatively analyzing four inhibitors (i.e., EDTA, EGTA, histidine and clioquinol) of Cu(2+)/Zn(2+) induced Aβ assembly. The inhibition efficiencies (e.g., half maximal inhibitory concentration, IC50 value) of these inhibitors could be measured in this work. As far as we know, these IC50 values were reported at the first time. In this assay, the streptavidin conjugated GNPs (SA-GNPs) were employed as indicators to monitor the Cu(2+)/Zn(2+) induced aggregating/disaggregating behaviors of biotin modified β-amyloid 1-16 peptides (Aβ1-16(biotin)). Because of high affinity of streptavidin (SA) with biotin, the aggregating/disaggregating of Aβ1-16(biotin) results in the significant color change of SA-GNPs. Furthermore, we demonstrate that the assay can be used as an effective tool for designing anti-dementia drugs through quantitative analysis of the interactions of four representative inhibitors with Cu(2+)/Zn(2+) induced Aβ assembly.

Topics: Affinity Labels; Amyloid beta-Peptides; Biotin; Chlorides; Circular Dichroism; Clioquinol; Colorimetry; Copper Sulfate; Edetic Acid; Egtazic Acid; Gold; Histidine; Inhibitory Concentration 50; Metal Nanoparticles; Microscopy, Electron, Transmission; Peptide Fragments; Protein Aggregates; Streptavidin; Zinc Compounds

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 (AbetaP) 1-40 into lipid membranes. Based on the conductance, kinetics, selectivity, and pharmacological properties, the following AbetaP[1-40]-formed ion channels have been identified: (i) The AbetaP[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 KCl cis/trans) at -160 mV, and the nonlinear current-voltage relationship drawn to a third-order polynomial, (b) selectivity sequence PK > PNa > PLi = 1.0:0.60:0.47, (c) Po 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 AbetaP[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 KCl) at +140 mV 17 pS (250/50 KCl) at -160 mV, the current-voltage relationship nonlinear drawn to a third-order polynomial fit, and (c) selectivity sequence PRb > (iii) The AbetaP[1-40]-formed medium conductance channel was charcterized 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 Vms more negative than -120 and more positive than +120 mV. (iv) The AbetaP[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 KCI) at +140 mV and 0-704 pS (in 250/50 mM KCl) 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 PK/Pcholine for the fast mode was 3.9 and selectivity sequence PK > PCs > PNa > PLi = 1.0:0.94:0.87:0.59. The value of PK/Pcholine for the slow mode was 2.7 and selectivity sequence PK > FNa > PLi > PCs = 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 mechani

Topics: Alzheimer Disease; Amyloid beta-Peptides; Cations; Cesium; Chlorides; Humans; Ion Channel Gating; Ion Channels; Kinetics; Lipid Bilayers; Lithium Chloride; Membrane Potentials; Patch-Clamp Techniques; Peptide Fragments; Potassium Chloride; Rubidium; Signal Transduction; Sodium Chloride; Structure-Activity Relationship; Zinc Compounds