Zinc binding to amyloid-beta: isothermal titration calorimetry and Zn competition experiments with Zn sensors.
Aggregation of the peptide amyloid-beta (Abeta) to amyloid plaques is a key event in Alzheimer's disease. According to the amyloid cascade hypothesis, Abeta aggregates are toxic to neurons via the production of reactive oxygen species and are hence directly involved in the cause of the disease. Zinc ions play an important role, because they are able to bind to Abeta and influence the aggregation properties. In the present work isothermal titration calorimetry and Zn sensors (zincon, Newport Green, and zinquin) were used to investigate the interaction of Zn with the full-length Abeta1-40 and Abeta1-42, as well as the truncated Abeta1-16 and Abeta1-28. The results suggest that Zn binding to Abeta induces a release of approximately 0.9 proton by the peptide. This correspond to the expected value upon Zn binding to the three histidines and indicates that further ligands are not deprotonated upon Zn binding. Such behavior is expected for carboxylates, but not the N-terminus. Moreover, the apparent dissociation constant (Kd,app) of Zn binding to all forms of Abeta is in the low micromolar range (1-20 microM) and rather independent of the aggregation state including soluble Abeta, Abeta fibrils, or Zn-induced Abeta aggregates. Finally, Zn in the soluble or aggregated Zn-Abeta form is well accessible for Zn chelators. The potential repercussions on metal chelation therapy are discussed.
Talmard
Christine
C
Laboratoire de Chimie de Coordination, CNRS UPR 8241, University Toulouse III, 205 route de Narbonne, 31077 Toulouse Cedex 4, France.
Bouzan
Anaïs
A
Faller
Peter
P
D000544:Alzheimer Disease / Q000378:metabolism; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D002151:Calorimetry / Q000379:methods; D007700:Kinetics; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D013816:Thermodynamics; D015032:Zinc / Q000737:chemistry / Q000378:metabolism*
D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; C433241:amyloid beta-protein (1-16); C084358:amyloid beta-protein (1-28); C078592:amyloid beta-protein (1-40); D015032:Zinc