Highly efficient soluble expression, purification and characterization of recombinant Aβ42 from Escherichia coli .

Aggregation of amyloid-β protein (Aβ) is hypothesized to be a seminal neuropathological event in Alzheimer's disease (AD). Recombinant expression and purification of Aβ represents a common basis for investigating the molecular mechanisms of amyloid formation and toxicity. Herein, we report a novel high-yield expression and purification method for Aβ42 based on fusion with maltose binding protein (MBP) followed by the soluble polypeptide linker (NANP) 3 and a modified tobacco etch virus (TEV) cleavage site before the Aβ42. We obtained a final yield of ∼18 mg L -1 of recombinant Aβ42 that was confirmed by SDS-PAGE, protein immunoblotting and MALDI-TOF. Finally, thioflavin T fluorescence and atomic force microscopy revealed that the recombinant Aβ42 aggregated into long, branched fibrils. Furthermore, the aggregates of the recombinant peptide had a strong cytotoxic effect on PC12 cells. The method described here can therefore be used to efficiently express the soluble fusion protein MBP-Aβ42 and obtain high-purity Aβ42 peptide, which can be used to understand the molecular mechanism of Aβ42 fibrillization and screen new candidate drugs for AD.