Abstract
The formation of amyloid-β peptide (Aβ) oligomers at the cellular membrane is considered a crucial process that underlies neurotoxicity in Alzheimer’s disease (AD). To obtain structural information on this type of oligomers, we were inspired by membrane protein approaches used to stabilize, characterize, and analyze the function of such proteins. Using these approaches, we developed conditions under which Aβ42, the Aβ variant most strongly linked to the aetiology of AD, assembles into an oligomer that inserts into lipid bilayers as a well-defined pore and adopts a specific structure with characteristics of a β-barrel arrangement. We named this oligomer β-barrel Pore-Forming Aβ42 Oligomer (βPFOAβ42). Here, we describe detailed protocols for its preparation and characterization. We expect βPFOAβ42 to be useful in establishing the involvement of membrane-associated Aβ oligomers in AD.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Arispe N, Rojas E, Pollard HB (1993) Alzheimer disease amyloid beta protein forms calcium channels in bilayer membranes: blockade by tromethamine and aluminum. Proc Natl Acad Sci U S A 90:567–571
Arispe N, Pollard HB, Rojas E (1996) Zn2+ interaction with Alzheimer amyloid beta protein calcium channels. Proc Natl Acad Sci U S A 93:1710–1715
Hirakura Y, Lin MC, Kagan BL (1999) Alzheimer amyloid Aβ1-42 channels: effects of solvent, pH, and congo red. J Neurosci Res 57:458–466
Lin H, Bhatia R, Lal R (2001) Amyloid beta protein forms ion channels: implications for Alzheimer’s disease pathophysiology. FASEB J 15:2433–2444
Kourie JI, Henry CL, Farrelly P (2001) Diversity of amyloid beta protein fragment [1-40]-formed channels. Cell Mol Neurobiol 21:255–284
Kayed R, Sokolov Y, Edmonds B et al (2004) Permeabilization of lipid bilayers is a common conformation-dependent activity of soluble amyloid oligomers in protein misfolding diseases. J Biol Chem 279:46363–46366
Quist A, Doudevski I, Lin H et al (2005) Amyloid ion channels: a common structural link for protein-misfolding disease. Proc Natl Acad Sci U S A 102:1–6
Laurén J, Gimbel DA, Nygaard HB et al (2009) Cellular prion protein mediates impairment of synaptic plasticity by amyloid-beta oligomers. Nature 457:1128–1132
Bode DC, Baker MD, Viles JH (2017) Ion channel formation by amyloid-β42 oligomers but not amyloid-β40 in cellular membranes. J Biol Chem 292:1404–1413
Popot J-L (2010) Amphipols, nanodiscs, and fluorinated surfactants: three nonconventional approaches to studying membrane proteins in aqueous solutions. Annu Rev Biochem 79:737–775
Sanders CR, Sönnichsen F (2006) Solution NMR of membrane proteins: practice and challenges. Magn Reson Chem 44:S24–S40
Columbus L, Lipfert J, Jambunathan K et al (2009) Mixing and matching detergents for membrane protein NMR structure determination. J Am Chem Soc 131:7320–7326
Hiller S, Garces RG, Malia TJ et al (2008) Solution structure of the integral human membrane protein VDAC-1 in detergent micelles. Science 321:1206–1210
Barrett PJ, Song Y, Van Horn WD et al (2012) The Amyloid Precursor Protein has a flexible transmembrane domain and binds cholesterol. Science 336:1168–1171
Laganowsky A, Reading E, Hopper JTS, Robinson CV (2013) Mass spectrometry of intact membrane protein complexes. Nat Protoc 8:639–651
Inoue S (2008) In situ Abeta pores in AD brain are cylindrical assembly of Abeta protofilaments. Amyloid 15:223–233
Serra-Batiste M, Ninot-Pedrosa M, Bayoumi M et al (2016) Aβ42 assembles into specific β-barrel pore-forming oligomers in membrane-mimicking environments. Proc Natl Acad Sci U S A 113:10866–10871
Religa TL, Sprangers R, Kay LE (2010) Dynamic regulation of archaeal proteasome gate opening as studied by TROSY NMR. Science 328:98–102
Shao H, Jao S-C, Ma K, Zagorski MG (1998) Solution structures of micelle-bound amyloid β-(1-40) and β-(1-42) peptides of Alzheimer’s disease. J Mol Biol 285:1–19
Teplow DB (2006) Preparation of amyloid β-protein for structural and functional studies. Methods Enzymol 413:20–33
Conibear AC, Daly NL, Craik DJ (2012) Quantification of small cyclic disulfide-rich peptides. Biopolymers 98:518–524
Tew DJ, Bottomley SP, Smith DP et al (2008) Stabilization of neurotoxic soluble beta-sheet-rich conformations of the Alzheimer’s disease amyloid-beta peptide. Biophys J 94:2752–2766
Turro NJ, Yekta A (1978) Luminescent probes for detergent solutions. A simple procedure for determination of the mean aggregation number of micelles. J Am Chem Soc 100:5951–5952
Tugarinov V, Hwang PM, Ollerenshaw JE, Kay LE (2003) Cross-correlated relaxation enhanced 1H-13C NMR spectroscopy of methyl groups in very high molecular weight proteins and protein complexes. J Am Chem Soc 125:10420–10428
Acknowledgment
M.N.P. and E.P. acknowledge the Spanish Government FPI program for predoctoral fellowships. This work was supported by Program Grants from the MINECO-FEDER (SAF2015-68789), from the Fundació La Marató de TV3 (20140730/31), from the Fondation pour la Researche Médicale (FRM)—Amorçage de Jeunes Equips (AJE20151234751), and from the Counseil Régional d’Aquitaine Limousin Poitou-Charentes (2016-1R30117) to N.C.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Serra-Batiste, M., Ninot-Pedrosa, M., Puig, E., Ciudad, S., Gairí, M., Carulla, N. (2018). Preparation of a Well-Defined and Stable β-Barrel Pore-Forming Aβ42 Oligomer. In: Sigurdsson, E., Calero, M., Gasset, M. (eds) Amyloid Proteins. Methods in Molecular Biology, vol 1779. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7816-8_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7816-8_2
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7815-1
Online ISBN: 978-1-4939-7816-8
eBook Packages: Springer Protocols