Disaggregation of Aβ42 for Structural and Biochemical Studies
The amyloid-β (Aβ) peptides that form the amyloid fibrils associated with Alzheimer’s disease are generated by sequential proteolysis of the amyloid precursor protein by β- and γ-secretase. The two predominant Aβ peptides, Aβ40 and Aβ42, differ by two amino acids, are soluble as monomers at low concentration (and/or low temperature) and are normally cleared from the brain parenchyma. In order to study the structure and assembly of these peptides, they are often synthesized using solid-phase peptide synthesis and purified. Here, we outline the method we use to prepare monomeric Aβ for structural and biochemical studies.
Key wordsAβ peptides Aβ40 Aβ42 Alzheimer’s disease FTIR NMR
This work was supported by NIH-NSF instrumentation grants (S10 RR13889 and DBI-9977553), a grant from the NIH to S.O.S (AG 27317). We gratefully acknowledge the W.M. Keck Foundation for support of the NMR facilities in the Center of Structural Biology at Stony Brook.
- 4.Portelius E, Bogdanovic N, Gustavsson M, Volkmann I, Brinkmalm G, Zetterberg H, Winblad B, Blennow K (2010) Mass spectrometric characterization of brain amyloid beta isoform signatures in familial and sporadic Alzheimer’s disease. Acta Neuropathol 120:185–193CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Pham E, Crews L, Ubhi K, Hansen L, Adame A, Cartier A, Salmon D, Galasko D, Michael S, Savas JN et al (2010) Progressive accumulation of amyloid-beta oligomers in Alzheimer’s disease and in amyloid precursor protein transgenic mice is accompanied by selective alterations in synaptic scaffold proteins. FEBS J 277:3051–3067CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Shankar GM, Leissring MA, Adame A, Sun XY, Spooner E, Masliah E, Selkoe DJ, Lemere CA, Walsh DM (2009) Biochemical and immunohistochemical analysis of an Alzheimer’s disease mouse model reveals the presence of multiple cerebral Aβ assembly forms throughout life. Neurobiol Dis 36:293–302CrossRefPubMedPubMedCentralGoogle Scholar
- 19.Broersen K, Jonckheere W, Rozenski J, Vandersteen A, Pauwels K, Pastore A, Rousseau F, Schymkowitz J (2011) A standardized and biocompatible preparation of aggregate-free amyloid beta peptide for biophysical and biological studies of Alzheimer’s disease. Protein Eng Des Sel 24:743–750CrossRefPubMedGoogle Scholar