Solution NMR Studies of the Aβ(1−40) and Aβ(1−42) Peptides Establish that the Met35 Oxidation State Affects the Mechanism of Amyloid Formation

• Published in: Journal of the American Chemical Society 2004-02, Vol.126 (7), p.1992-2005
• Main Authors: Hou, Liming, Shao, Haiyan, Zhang, Yongbo, Li, Hua, Menon, Nanda K, Neuhaus, Elizabeth B, Brewer, John M, Byeon, In-Ja L, Ray, Dale G, Vitek, Michael P, Iwashita, Takashi, Makula, Ronald A, Przybyla, Alan B, Zagorski, Michael G
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amyloid - biosynthesis; Amyloid beta-Peptides - chemistry; Amyloid beta-Peptides - metabolism; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Hydrophobic and Hydrophilic Interactions; Interactions. Associations; Intermolecular phenomena; Methionine - chemistry; Methionine - metabolism; Models, Molecular; Molecular biophysics; Molecular Sequence Data; Nuclear Magnetic Resonance, Biomolecular - methods; Oxidation-Reduction; Peptide Fragments - chemistry; Peptide Fragments - metabolism; Protein Structure, Secondary; Solutions
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja036813f

The pathogenesis of Alzheimer's disease is characterized by the aggregation and fibrillation of the 40-residue Aβ(1−40) and 42-residue Aβ(1−42) peptides into amyloid plaques. The structural changes associated with the conversion of monomeric Aβ peptide building blocks into multimeric fibrillar β-strand aggregates remain unknown. Recently, we established that oxidation of the methionine-35 side chain to the sulfoxide (Met35red → Met35ox) significantly impedes the rate of aggregation and fibrillation of the Aβ peptide. To explore this effect at greater resolution, we carefully compared the 1H, 15N, and 13C NMR chemical shifts of four Aβ peptides that had the Met35 reduced or oxidized (Aβ(1−40)Met35red, Aβ(1−40)Met35ox, Aβ(1−42)Met35red, and Aβ(1−42)Met35ox). With the use of a special disaggregation protocol, the highly aggregation prone Aβ peptides could be studied at higher, millimolar concentrations (as required by NMR) in aqueous solution at neutral pH, remaining largely monomeric at 5 °C as determined by sedimentation equilibrium studies. The NOE, amide-NH temperature coefficients, and chemical shift indices of the 1Hα, 13Cα, and 13Cβ established that the four peptides are largely random, extended chain structures, with the Met35ox reducing the propensity for β-strand structure at two hydrophobic regions (Leu17−Ala21 and Ile31−Val36), and turn- or bendlike structures at Asp7−Glu11 and Phe20−Ser26. Additional NMR studies monitoring changes that occur during aging at 37 °C established that, along with a gradual loss of signal/noise, the Met35ox significantly hindered upfield chemical shift movements of the 2H NMR signals for the His6, His13, and His14 side chains. Taken together, the present NMR studies demonstrate that the Met35red → Met35ox conversion prevents aggregation by reducing both hydrophobic and electrostatic association and that the Aβ(1−40)Met35red, Aβ(1−40)Met35ox, Aβ(1−42)Met35red, and Aβ(1−42)Met35ox peptides may associate differently, through specific, sharp changes in structure during the initial stages of aggregation.