title: Preparation of Amyloid Fibrils for Magic-Angle Spinning Solid-State NMR Spectroscopy.
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.
authors.affiliation: Department of Chemistry, University of Illinois at Urbana-Champaign, A129B CLSL, Box 50-6, 600 South Mathews Avenue, Urbana, IL, 61801, USA.
authors.forename: Marcus D
authors.initials: MD
authors.lastname: Tuttle
authors.affiliation: Department of Chemistry, University of Illinois at Urbana-Champaign, A129B CLSL, Box 50-6, 600 South Mathews Avenue, Urbana, IL, 61801, USA.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Courtney
authors.affiliation: Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Barclay
authors.affiliation: Department of Chemistry, University of Illinois at Urbana-Champaign, A129B CLSL, Box 50-6, 600 South Mathews Avenue, Urbana, IL, 61801, USA. rienstra@illinois.edu.|Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA. rienstra@illinois.edu.|Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA. rienstra@illinois.edu.
authors.forename: Chad M
authors.initials: CM
authors.lastname: Rienstra
mesh_terms: D000682:Amyloid / Q000737:chemistry* / Q000302:isolation & purification; D058227:Amyloidogenic Proteins / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011487:Protein Conformation; D051844:alpha-Synuclein / Q000737:chemistry / Q000235:genetics
chemical_list: D000682:Amyloid; D058227:Amyloidogenic Proteins; D051844:alpha-Synuclein
keywords: Amyloid fibrils; Fibrillization; Size exclusion chromatography; Solid-state NMR; α-synuclein