Metal-binding properties of the peptide APP170-188: a model of the ZnII-binding site of amyloid precursor protein (APP).
Amyloid precursor protein (APP) plays a key role in Alzheimer's disease (AD), although the function of this membrane protein is still unclear. Metal ions are implicated in AD and they also interact with APP. APP possesses a strong ZnII binding site, which is evolutionary conserved. In this paper a synthetic peptide, APP170-188, with a sequence corresponding to the conserved ZnII-binding domain of APP, was synthesised and its metal-binding properties analysed. Titration experiments pointed to the binding of a stoichiometric amount of divalent ions. Further studies indicated that the binding of divalent metals like ZnII, CdII and CoII induces the dimerisation of the peptide. This dimer contains a dinuclear cluster in which the two divalent metals are bridged by two thiolate ligands from cysteine residues. The other two ligands of the tetrahedral coordination sites of each metal ion are terminal thiolate ligands. This structure was supported by the following arguments. The complex formed with CoII presents the characteristic features for tetrahedral tetrathiolate coordination in its UV-visible spectrum. The sequence of APP170-188 contains only three cysteine residues, which is incompatible with a monomeric CoII-APP170-188 complex. EPR measurements of the complex with one equivalent of CoII show almost no signal at 4 K, which is compatible with an antiferromagnetic spin-coupling of the metal ions in a cluster structure. Size-exclusion chromatography indicated that the elution time for the complexes with ZnII and CdII corresponds to the expected molecular weight of a dimer. The circular dichroism (CD) spectrum of the complex with one equivalent of CdII shows a band at 265 nm+, and an ellipticity similar to those observed for similar CdII-thiolate clusters. Possible biological implications of the ZnII binding site and the metal-induced dimerisation are discussed.
Ciuculescu
Eliza-Diana
ED
Laboratoire de Chimie de Coordination, CNRS UPR 8241 (associated with University Toulouse III), 205 route de Narbonne, 31077 Toulouse Cedex, France.
Mekmouche
Yasmina
Y
Faller
Peter
P
D016564:Amyloid beta-Protein Precursor / Q000737:chemistry*; D001665:Binding Sites; D002104:Cadmium / Q000737:chemistry*; D003035:Cobalt / Q000737:chemistry*; D008024:Ligands; D008956:Models, Chemical; D009942:Organometallic Compounds / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D013329:Structure-Activity Relationship; D015032:Zinc / Q000737:chemistry*
C511644:APP170-188 peptide; D016564:Amyloid beta-Protein Precursor; D008024:Ligands; D009942:Organometallic Compounds; D010446:Peptide Fragments; D002104:Cadmium; D003035:Cobalt; D015032:Zinc