Solution 1H NMR investigation of Zn2+ and Cd2+ binding to amyloid-beta peptide (Abeta) of Alzheimer's disease.
Elevated levels of zinc2+ and copper2+ are found chelated to the amyloid-beta-peptide (Abeta) in isolated senile plaque cores of Alzheimer's disease (AD) patients. However, the precise residues involved in Zn2+ ligation are yet to be established. We have used 1H NMR and CD to probe the binding of Zn2+ to Abeta(1-28). Zinc binding to Abeta causes a number of 1H NMR resonances to exhibit intermediate exchange broadening upon Zn2+ addition, signals in slow and fast exchange are also observed. In addition, there is a general loss of signal for all resonances with Zn2+ addition, suggestive of the formation of high molecular weight polymeric species. Perturbations in specific 1H NMR resonances between residues 6 and 14, and analysis of various Abeta analogues in which each of the three His residues have been replaced by alanine, indicates that His6, His13 and His14 residues are implicated in Zn-Abeta binding. Complementary studies with Cd2+ ions cause perturbations to 1H NMR spectra that are strikingly similar to that observed for Zn2+. Binding monitored at Val12 indicates a 1:1 stoichiometry with Abeta for both Zn2+ and Cd2+ ions. Circular Dichroism (CD) studies in the far-UV indicate quite minimal ordering of the main-chain with Zn2+ or Cd2+ addition. Changes in the far-UV are quite different from that obtained with Cu2+ additions indicating that Zn2+ coordination is distinct from that of Cu2+ ions. Taken together, these observations seem to suggest that Zn2+ coordination is dominated by inter-molecular coordination and the formation of polymeric species.
Syme
Christopher D
CD
School of Biological Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, UK.
Viles
John H
JH
D000544:Alzheimer Disease / Q000378:metabolism; D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000235:genetics; D002104:Cadmium / Q000737:chemistry*; D002942:Circular Dichroism; D006801:Humans; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D012996:Solutions / Q000737:chemistry; D015032:Zinc / Q000737:chemistry*
D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; D010455:Peptides; D012996:Solutions; C433241:amyloid beta-protein (1-16); C084358:amyloid beta-protein (1-28); D002104:Cadmium; D015032:Zinc