Preparation of Amyloid Fibrils for Magic-Angle Spinning Solid-State NMR Spectroscopy

Methods Mol Biol. 2016:1345:173-83. doi: 10.1007/978-1-4939-2978-8_11.

Abstract

Solid-state NMR spectroscopy (SSNMR) is an established and invaluable tool for the study of amyloid fibril structure with atomic-level detail. Optimization of the homogeneity and concentration of fibrils enhances the resolution and sensitivity of SSNMR spectra. Here, we present a fibrillization and fibril processing protocol, starting from purified monomeric α-synuclein, that enables the collection of high-resolution SSNMR spectra suitable for site-specific structural analysis. This protocol does not rely on any special features of α-synuclein and should be generalizable to any other amyloid protein.

Keywords: Amyloid fibrils; Fibrillization; Size exclusion chromatography; Solid-state NMR; α-synuclein.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amyloid / chemistry*
  • Amyloid / isolation & purification
  • Amyloidogenic Proteins / chemistry*
  • Amyloidogenic Proteins / isolation & purification
  • Humans
  • Magnetic Resonance Spectroscopy / methods*
  • Protein Conformation
  • alpha-Synuclein / chemistry
  • alpha-Synuclein / genetics

Substances

  • Amyloid
  • Amyloidogenic Proteins
  • alpha-Synuclein