Observation of Intact and Proteolytically Cleaved Amyloid-Beta (1-40)-Oleuropein Noncovalent Complex at Neutral pH by Mass Spectrometry

Mass spectrometry analyses carried out on mass spectrometers equipped with soft ionization sources demonstrated their utility in the assessment of the formation of noncovalent complexes and the localization of the binding sites. Direct analyses by mass spectrometry of the noncovalent complex formed in acidic and mildly acidic environments by amyloid beta (1-40) peptide and oleuropein have been previously described, and, in several studies, the absorption, metabolism, excretion, and the implications in the prevention and therapy of Alzheimer's disease of oleuropein have been investigated. Our paper presents modifications of the method previously employed for noncovalent complex observation, namely, the amyloid beta (1-40) pretreatment, followed by an increase in the pH and replacement of the chemical environment from ammonium acetate to ammonium bicarbonate. The formation of noncovalent complexes with one or two molecules of oleuropein was detected in all chemical solutions used, and the amyloid beta (17-28) binding site was identified via proteolytic experiments using trypsin prior to and after noncovalent complex formation. Our results highlight the importance of further studies on the effect of oleuropein against amyloid beta aggregation.

Automated summary

This study used mass spectrometry to investigate the formation of noncovalent complexes between oleuropein and amyloid beta. By modifying the chemical environment and conducting proteolytic experiments, the binding site was identified. The results highlight the importance of further research on the effect of oleuropein against amyloid beta aggregation.