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NMR Reveals Anomalous Copper(II) Binding to the Amyloid Aβ Peptide of Alzheimer's Disease

2006; American Chemical Society; Volume: 128; Issue: 29 Linguagem: Inglês

10.1021/ja046032u

1943-2984

Liming Hou, Michael G. Zagorski,

Drug Transport and Resistance Mechanisms

The Aβ peptide is the major protein component of amyloid deposits in Alzheimer's disease (AD). Age-related microenvironmental changes in the AD brain promote amyloid formation that leads to cell injury and death. Altered levels of metals (such as Cu and Zn) exist in the AD brain, and because Cu and Zn can be bound to the Aβ in the amyloid plaques, it is thought that these binding events in vivo may trigger or prevent Aβ amyloid formation in the AD brain. Although several structural models have been proposed, all of these are undefined due to the lack of definitive structural data. The present NMR studies utilized uniformly 15N-labeled Aβ(1−40) peptide and 1H−15N HSQC experiments and demonstrate for the first time that the Aβ binds Cu and Zn in a distinct manner. The binding promotes NH signal disappearance of E3-V18, which was not due to the paramagnetic effect of Cu2+, as identical NMR studies were seen with Zn2+, which is diamagnetic. NMR titration experiments showed that the amide NH peak intensities of R5-L17 showed the most pronounced intensity reduction, and that the 1H signals for the side chain aromatic signals of the three histidines shift upfield (H6, H13, and H14). We propose that initially Cu2+ is anchored to the Aβ monomer (fast exchange rate) and is followed by deprotonation and/or severe line broadening of the backbone amide NH for E3-V18 (intermediate exchange rate). By contrast, Cu2+ binding to soluble Aβ aggregates leads to rapid aggregation and nonfibrillar amorphous structures, and without metal, the Aβ can undergo the normal time-dependent aggregation, eventually producing more ordered, late-stage parallel β-sheet structures. These anomalous (rare) binding events may account for some of the unique properties associated with the Aβ, such as its proposed "dual role", where sequestration of metal ions by the monomer is neuroprotective, while that by β-aggregates generates oxygen radicals and causes neuronal death.