Open Access
Higher Order Amyloid Fibril Structure by MAS NMR and DNP Spectroscopy
Galia T. Debelouchina,Marvin J. Bayro,Anthony W. P. Fitzpatrick,Vladimir Ladizhansky,Michael T. Colvin,Marc A. Caporini,Christopher P. Jaroniec,Vikram S. Bajaj,Melanie Rosay,Cait E. MacPhee,Michele Vendruscolo,Werner Maas,Christopher M. Dobson,Robert G. Griffin +13 more
TLDR
In this paper, a protein magic angle spinning (MAS) NMR spectroscopy has been applied to the fibrils formed by an eleven residue segment of the amyloidogenic protein transthyretin.Abstract:
Protein magic angle spinning (MAS) NMR spectroscopy has generated structural models of several amyloid fibril systems, thus providing valuable information regarding the forces and interactions that confer the extraordinary stability of the amyloid architecture. Despite these advances, however, obtaining atomic resolution information describing the higher levels of structural organization within the fibrils remains a significant challenge. Here, we detail MAS NMR experiments and sample labeling schemes designed specifically to probe such higher order amyloid structure, and we have applied them to the fibrils formed by an eleven-residue segment of the amyloidogenic protein transthyretin (TTR(105−115)). These experiments have allowed us to define unambiguously not only the arrangement of the peptide β-strands into β-sheets but also the β-sheet interfaces within each protofilament, and in addition to identify the nature of the protofilament-to-protofilament contacts that lead to the formation of the complete fibril. Our efforts have resulted in 111 quantitative distance and torsion angle restraints (10 per residue) that describe the various levels of structure organization. The experiments benefited extensively from the use of dynamic nuclear polarization (DNP), which in some cases allowed us to shorten the data acquisition time from days to hours and to improve significantly the signal-to-noise ratios of the spectra. The β-sheet interface and protofilament interactions identified here revealed local variations in the structure that result in multiple peaks for the exposed N- and C-termini of the peptide and in inhomogeneous line-broadening for the residues buried within the interior of the fibrils.read more
Citations
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疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A
TL;DR: PfPMP1)与感染红细胞、树突状组胞以及胎盘的单个或多个受体作用,在黏附及免疫逃避中起关键的作�ly.
Journal ArticleDOI
Atomic Resolution Structure of Monomorphic Aβ42 Amyloid Fibrils
Michael T. Colvin,Robert Silvers,Qing Zhe Ni,Thach V. Can,Ivan V. Sergeyev,Melanie Rosay,Kevin J. Donovan,Brian Michael,Joseph S. Wall,Sara Linse,Robert G. Griffin +10 more
TL;DR: The structure provides a point of departure for the design of drugs that bind to the fibril surface and therefore interfere with secondary nucleation and for other therapeutic approaches to mitigate Aβ42 aggregation.
Journal ArticleDOI
Dynamic nuclear polarization for sensitivity enhancement in modern solid-state NMR.
TL;DR: This review provides an in-depth overview of the relevant topics involved in DNP-enhanced MAS NMR spectroscopy and underlines that MAS DNP has moved far beyond the proof-of-concept stage and has become an important tool for research in these fields.
Journal ArticleDOI
Nuclear depolarization and absolute sensitivity in magic-angle spinning cross effect dynamic nuclear polarization
Frederic Mentink-Vigier,Subhradip Paul,Daniel Lee,Akiva Feintuch,Sabine Hediger,Sabine Hediger,Shimon Vega,Gaël De Paëpe +7 more
TL;DR: It is demonstrated that TOTAPOL and AMUPOL both lead to observable depolarization at ≈110 K, and that the magnitude of this depolarized nuclear state is radical dependent.
Journal ArticleDOI
Magic Angle Spinning NMR of Proteins: High-Frequency Dynamic Nuclear Polarization and 1H Detection
TL;DR: High-frequency dynamic nuclear polarization (DNP) and (1)H-detected MAS techniques are discussed, which overcome NMR's inherent low sensitivity and permits structural investigations at the nanomolar scale.
References
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疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A
TL;DR: PfPMP1)与感染红细胞、树突状组胞以及胎盘的单个或多个受体作用,在黏附及免疫逃避中起关键的作�ly.
Journal ArticleDOI
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Heteronuclear decoupling in rotating solids
Andrew E. Bennett,Chad M. Rienstra,Michèle Auger,Michèle Auger,K. V. Lakshmi,K. V. Lakshmi,Robert G. Griffin +6 more
TL;DR: In this article, a simple two pulse phase modulation (TPPM) scheme was proposed to reduce the residual linewidths arising from insufficient proton decoupling power in double resonance magic angle spinning (MAS) experiments.