Self-association of β-Amyloid Peptide (1–40) in Solution and Binding to Lipid Membranes

Abstract

The β-amyloid peptide (βAP), a 39 to 43 residue peptide, is the major component of Alzheimer plaques. Using circular dichroism spectroscopy, titration calorimetry, and analytical, ultracentrifugation we have analyzed the self-association of βAP(1–40) in aqueous solution and the binding of βAP(1–40) to negatively charged lipid vesicles. The CD spectra of both aggregation and membrane binding are characterized by an iso-dichroic point at 212 nm, indicating a simple two-state equilibrium for both cases. In aqueous solution βAP(1–40) exhibits a reversible, concentration-dependent random coil ⇄ β-structure transition which can be described by a cooperative aggregation model with an association constant ofs= 1.05 x 104M−1and a nucleation parameter of σ = 0.012. A similar conformational change is observed upon addition of lipid. At a given peptide concentration, the addition of negatively charged, small unilamellar vesicles also induces a conformational change from a random coil conformation to a conformation with 40 to 60% β-structure. The binding isotherm can be measured with high sensitivity titration calorimetry. It is approximately linear in the initial binding phase and exhibits an apparent saturation behaviour. The apparent binding constant decreases with concentration from Kapp≈ 2100 M−1at low concentration to 700 M−1at the highest concentration measured. Peptide penetration into the lipid membrane and peptide aggregation at the membrane surface are proposed as possible mechanisms to explain the lipid-induced random coil ⇄ β-structure transition.