Characterization of 15N Chemical Shift and 1H−15N Dipolar Coupling Interactions in a Peptide Bond of Uniaxially Oriented and Polycrystalline Samples by One-Dimensional Dipolar Chemical Shift Solid-State NMR Spectroscopy

Lee, D. K.; Wittebort, Richard J.; Ramamoorthy, Ayyalusamy

doi:10.1021/ja981599u

Cited by 64 publications

(107 citation statements)

References 62 publications

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“…For instance, Lee et (98). The experiment can be used to more accurately define the orientation of the CSA tensor with respect to the NÀ ÀH bond and, by fitting spectral simulations, the orientation of the NÀ ÀH bond with respect to the fiber axis can be determined (97).…”

Section: Measurement Of Interaction Tensors In Oriented Samplesmentioning

confidence: 99%

Studying natural structural protein fibers by solid‐state nuclear magnetic resonance

Arnold

Marcotte

2009

Concepts Magnetic Resonance

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As a consequence of evolutionary pressure, various organisms have developed structural fibers displaying a range of exceptional mechanical properties adapted specifically to their functions. An understanding of these properties at the molecular level requires a detailed description of local structure, orientation with respect to the fiber and size of constitutive units, and dynamics on various timescales. The size and lack of long-range order in these protein systems constitute an important challenge to classical structural techniques such as high-resolution NMR and X-ray diffraction. Solidstate NMR overcomes these constraints and is uniquely suited to the study of these inherently disordered systems. Solid-state NMR experiments developed or applied to determine structure, orientation, and dynamics of these complex proteins will be reviewed and illustrated through examples of their applications to fibers such as spider and silkworm silks, collagen, elastin, and keratin.

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Section: Measurement Of Interaction Tensors In Oriented Samplesmentioning

confidence: 99%

Studying natural structural protein fibers by solid‐state nuclear magnetic resonance

Arnold

Marcotte

2009

Concepts Magnetic Resonance

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“…Errors on the reported tilt angles are estimated from the line widths observed in the chemical shift and dipolar coupling spectra. However, there could be additional contributions to the error from the variation of chemical shift tensors (57,64) from the model tensor used for these calculations and due to peptide dynamics that would have influenced the observed experimental parameters. Nevertheless, the reported tilt angles are in good agreement with results obtained from MD simulations, discussed below.…”

Section: Orientation Of Skp and Sa Peptides In Bilayersmentioning

confidence: 99%

Structure, Topology, and Tilt of Cell-Signaling Peptides Containing Nuclear Localization Sequences in Membrane Bilayers Determined by Solid-State NMR and Molecular Dynamics Simulation Studies

et al. 2007

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Cell-signaling peptides have been extensively used to transport functional molecules across the plasma membrane into living cells. These peptides consist of a hydrophobic sequence and a cationic nuclear localization sequence (NLS). It has been assumed that the hydrophobic region penetrates through the hydrophobic lipid bilayer and delivers the NLS inside the cell. To better understand the transport mechanism of these peptides, in this study, we investigated the structure, orientation, tilt of the peptide relative to the bilayer normal, and the membraneinteraction of two cell-signaling peptides, SA and SKP. Results from CD and solid-state NMR experiments combined with molecular dynamics simulations suggest that the hydrophobic region is helical and has a transmembrane orientation with the helical axis tilted away from the bilayer normal. The influence of the hydrophobic mismatch, between the hydrophobic length of the peptide and the hydrophobic thickness of the bilayer, on the tilt angle of the peptides was investigated using thicker POPC and thinner DMPC bilayers. NMR experiments showed that the hydrophobic domain of each peptide has a tilt angle of 15±3° in POPC, while in DMPC 25±3° and 30±3° tilts were observed for SA and SKP peptides respectively. These results are in good agreement with molecular dynamics simulations, which predicts a tilt angle of 13.3° (SA in POPC), 16.4° (SKP in POPC), 22.3° (SA in DMPC) and 31.7°( SKP in POPC). These results and simulations on the hydrophobic fragment of SA or SKP suggest that the tilt of helices increases with a decrease in the bilayer thickness without changing the phase, order, and structure of the lipid bilayers. KeywordsCell-permeable peptides; NLS; PISA wheel; tilt; hydrophobic mismatch; transmembrane helix Noninvasive delivery of peptides, proteins, oligonucleotides and other functional cargo molecules into living cells is highly dependent on their efficient transport across the plasma membrane barrier (1-7). Hydrophobic peptides have been successfully used to transport nuclear localization sequences (NLS) in order to probe intracellular signaling, which involved

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“…[1][2][3][4][5][6][7][8][9][10][11][12][13] Even though select non-isotropic properties of chemical shift tensors can be obtained through solution-state NMR approaches, solid-state NMR spectroscopy is far more suitable for deriving the anisotropic information. 14 In solids, CSA can be directly measured by recording powder patterns in static powder samples, but these experiments suffer from poor sensitivity, spectral overlap when multiple chemical sites are present, and from broadening and distortion to the CSA line shapes introduced by a) Author to whom correspondence should be addressed.…”

Section: Introductionmentioning

confidence: 99%

Recoupling of chemical shift anisotropy by R-symmetry sequences in magic angle spinning NMR spectroscopy

Hou

Byeon

Ahn

et al. 2012

The Journal of Chemical Physics

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C and15 N chemical shift (CS) interaction is a sensitive probe of structure and dynamics in a wide variety of biological and inorganic systems, and in the recent years several magic angle spinning NMR approaches have emerged for residue-specific measurements of chemical shift anisotropy (CSA) tensors in uniformly and sparsely enriched proteins. All of the currently existing methods are applicable to slow and moderate magic angle spinning (MAS) regime, i.e., MAS frequencies below 20 kHz. With the advent of fast and ultrafast MAS probes capable of spinning frequencies of 40-100 kHz, and with the superior resolution and sensitivity attained at such high frequencies, development of CSA recoupling techniques working under such conditions is necessary. In this work, we present a family of R-symmetry based pulse sequences for recoupling of 13 C/ 15 N CSA interactions that work well in both natural abundance and isotopically enriched systems. We demonstrate that efficient recoupling of either first-rank (σ 1 ) or second-rank (σ 2 ) spatial components of CSA interaction is attained with appropriately chosen γ -encoded RN n v symmetry sequences. The advantage of these γ -encoded RN n v -symmetry based CSA (RNCSA) recoupling schemes is that they are suitable for CSA recoupling under a wide range of MAS frequencies, including fast MAS regime. Comprehensive analysis of the recoupling properties of these RN n v symmetry sequences reveals that the σ 1 -CSA recoupling symmetry sequences exhibit large scaling factors; however, the partial homonuclear dipolar Hamiltonian components are symmetry allowed, which makes this family of sequences suitable for CSA measurements in systems with weak homonuclear dipolar interactions. On the other hand, the γ -encoded symmetry sequences for σ 2 -CSA recoupling have smaller scaling factors but they efficiently suppress the homonuclear dipole-dipole interactions. Therefore, the latter family of sequences is applicable for measurements of CSA parameters in systems with strong homonuclear dipolar couplings, such as uniformly-13 C labeled biological solids. We demonstrate RNCSA NMR experiments and numerical simulations establishing the utility of this approach to the measurements of 13

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Characterization of ¹⁵N Chemical Shift and ¹H−¹⁵N Dipolar Coupling Interactions in a Peptide Bond of Uniaxially Oriented and Polycrystalline Samples by One-Dimensional Dipolar Chemical Shift Solid-State NMR Spectroscopy

Cited by 64 publications

References 62 publications

Studying natural structural protein fibers by solid‐state nuclear magnetic resonance

Studying natural structural protein fibers by solid‐state nuclear magnetic resonance

Structure, Topology, and Tilt of Cell-Signaling Peptides Containing Nuclear Localization Sequences in Membrane Bilayers Determined by Solid-State NMR and Molecular Dynamics Simulation Studies

Recoupling of chemical shift anisotropy by R-symmetry sequences in magic angle spinning NMR spectroscopy

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