Solid-State NMR Studies of Amyloid Materials: A Protocol to Define an Atomic Model of Aβ(1–42) in Amyloid Fibrils

  • Yiling Xiao
  • Dan McElheny
  • Minako Hoshi
  • Yoshitaka Ishii
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1777)

Abstract

Intense efforts have been made to understand the molecular structures of misfolded amyloid β (Aβ) in order to gain insight into the pathological mechanism of Alzheimer’s disease. Solid-state NMR spectroscopy (SSNMR) is considered a primary tool for elucidating the structures of insoluble and noncrystalline amyloid fibrils and other amyloid assemblies. In this chapter, we describe a detailed protocol to obtain the first atomic model of the 42-residue human Aβ peptide Aβ(1–42) in structurally homogeneous amyloid fibrils from our recent SSNMR study (Nat Struct Mol Biol 22:499–505, 2015). Despite great biological and clinical interest in Aβ(1–42) fibrils, their structural details have been long-elusive until this study. The protocol is divided into four sections. First, the solid-phase peptide synthesis (SPPS) and purification of monomeric Aβ(1–42) is described. We illustrate a controlled incubation method to prompt misfolding of Aβ(1–42) into homogeneous amyloid fibrils in an aqueous solution with fragmented Aβ(1–42) fibrils as seeds. Next, we detail analysis of Aβ(1–42) fibrils by SSNMR to obtain structural restraints. Finally, we describe methods to construct atomic models of Aβ(1–42) fibrils based on SSNMR results through two-stage molecular dynamics calculations.

Key words

Aβ(1–42) Amyloid fibrils Seeding method SSNMR Structure Atomic model 

Notes

Acknowledgments

This work was supported by the National Institutes of Health R01 and U01 programs (GM 098033) and Alzheimer’s Association IIRG grant (08-91256) for YI.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yiling Xiao
    • 1
  • Dan McElheny
    • 1
  • Minako Hoshi
    • 2
    • 3
  • Yoshitaka Ishii
    • 1
    • 4
  1. 1.Department of ChemistryUniversity of Illinois at ChicagoChicagoUSA
  2. 2.Institute of Biomedical Research and Innovation, FBRIKobeJapan
  3. 3.Department of Anatomy and Developmental Biology, Graduate School of MedicineKyoto UniversityKyotoJapan
  4. 4.School of Life Science and TechnologyTokyo Institute of TechnologyYokohamaJapan

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