Observation of spin exchange by two-dimensional fourier transform 13C cross polarization-magic-angle spinning

Szeverenyi, Nikolaus M.; Sullivan, Mark J.; Maciel, Gary E.

doi:10.1016/0022-2364(82)90213-x

Cited by 287 publications

(293 citation statements)

References 13 publications

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“…13 C chemical shifts were referenced externally relative to TMS. Two-dimensional 13 C-13 C proton-driven spin exchange spectra with a mixing time of 50 ms were acquired for peak assignment (21).…”

Section: Methodsmentioning

confidence: 99%

Dynamics of Amyloid β Fibrils Revealed by Solid-state NMR

Scheidt

Morgado

Rothemund

et al. 2012

Journal of Biological Chemistry

100

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Background: Alzheimer disease is the most important neurodegenerative disorder; treatment approaches require atomistic knowledge of fibrillar structure and dynamics. Results: We have site-specifically studied the molecular dynamics of amyloid ␤ (A␤) fibrils by solid-state NMR. Conclusion:The ␤-sheet motifs of A␤ are essentially rigid, and the termini exhibit more flexibility. Significance: Dynamics studies of A␤ fibrils suggest a structural role of the N terminus of the peptide.

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Section: Methodsmentioning

confidence: 99%

Dynamics of Amyloid β Fibrils Revealed by Solid-state NMR

Scheidt

Morgado

Rothemund

et al. 2012

Journal of Biological Chemistry

100

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“…Solid-state NMR spectroscopy All NMR-experiments were conducted using 3.2 mm tripleresonance ( 1 H, 13 C, 15 N) probe heads at static magnetic fields of 14.1 and 18.8 T (Agilent Technologies) and MAS frequencies of 11 kHz §3 Hz or 12.5 kHz §3 Hz, respectively, for ( 13 C, 13 C) PDSD- 71 and NCACX-/NCOCX-type spectra 72 or 23.0 kHz § 5 Hz for DREAM mixing, 73 respectively. Temperature was calibrated for each probe head and each spinning speed using nickelocene.…”

Section: Circular Dichroism (Cd) Spectroscopymentioning

confidence: 99%

Progress towards structural understanding of infectious sheep PrP-amyloid

Müller

Brener

Andréoletti

et al. 2014

Prion

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The still elusive structural difference of non-infectious and infectious amyloid of the mammalian prion protein (PrP) is a major pending milestone in understanding protein-mediated infectivity in neurodegenerative diseases. Preparations of PrP-amyloid proven to be infectious have never been investigated with a high-resolution technique. All available models to date have been based on low-resolution data. Here, we establish protocols for the preparation of infectious samples of full-length recombinant (rec) PrP-amyloid in NMR-sufficient amounts by spontaneous fibrillation and seeded fibril growth from brain extract. We link biological and structural data of infectious recPrP-amyloid, derived from bioassays, atomic force microscopy, and solid-state NMR spectroscopy. Our data indicate a semi-mobile N-terminus, some residues with secondary chemical shifts typical of a-helical secondary structure in the middle part between »115 to »155, and a distinct b-sheet core C-terminal of residue »155. These findings are not in agreement with all current models for PrP-amyloid. We also provide evidence that samples seeded from brain extract may not differ in the overall arrangement of secondary structure elements, but rather in the flexibility of protein segments outside the b-core region. Taken together, our protocols provide an essential basis for the high-resolution characterization of non-infectious and infectious PrP-amyloid in the near future.

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“…Field strengths for the spin lock pulses were 30 kHz on the 1 H and 13 kHz on the 13 C channel (with proton decoupling with a field strength of 76 kHz), respectively. For 2D 13 C/ 13 C correlation spectra, PDSD with mixing times from 20 to 300 ms (Bloembergen, 1949;Szeverenyi et al, 1982) was employed. DQ coherences for acquisition of DQS were excited and reconverted with the Supercycled POST-C5 (SPC5) sequence (Hohwy et al, 1999).…”

Section: Mas Solid-state Nmr Experimentsmentioning

confidence: 99%

Full-length Vpu and human CD4(372–433) in phospholipid bilayers as seen by magic angle spinning NMR

Quynh¹,

Wittlich²,

Glück³

et al. 2013

Biological Chemistry

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HIV-1 Vpu and CD4(372-433), a peptide comprising the transmembrane and cytoplasmic domain of human CD4, were recombinantly expressed in Escherichia coli, uniformly labeled with 13 C and 15 N isotopes, and separately reconstituted into phospholipid bilayers. Highly resolved dipolar cross-polarization (CP)-based solid-state NMR spectra of the two transmembrane proteins were recorded under magic angle sample spinning. Partial assignment of 13 C resonances was achieved. Site-specific assignments were obtained for 13 amino acid residues of CD4(372-433) and two Vpu residues. Additional amino acid type-specific assignments were achieved for 10 amino acid spin systems for both CD4(372-433) and Vpu. Further, structural flexibility was probed with different dipolar recoupling techniques, and the correct insertion of the transmembrane domains into the lipid bilayers was confirmed by proton spin diffusion experiments.

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Observation of spin exchange by two-dimensional fourier transform 13C cross polarization-magic-angle spinning

Cited by 287 publications

References 13 publications

Dynamics of Amyloid β Fibrils Revealed by Solid-state NMR

Dynamics of Amyloid β Fibrils Revealed by Solid-state NMR

Progress towards structural understanding of infectious sheep PrP-amyloid

Full-length Vpu and human CD4(372–433) in phospholipid bilayers as seen by magic angle spinning NMR

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