15N chemical shift and 15N-13C dipolar tensors for the peptide bond in [1-13C]glycyl[15N]glycine hydrochloride monohydrate

Harbison, Gerard S.; Jelinski, Lynn W.; Torchia, Dennis A.; Herzfeld, Judith; Griffin, Robert G.

doi:10.1016/0022-2364(84)90027-1

Cited by 75 publications

(266 citation statements)

References 11 publications

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“…A similar range of sitespecific 15 N CSA values (-129 to -213 ppm) was reported for ribonuclease H 8 , although with a somewhat different mean (-172 ppm), for a selection of well-ordered amides. For this subset of residues, the site-to-site variability in CSA was estimated to be ±5.5 ppm (upper limit ±9.6 ppm at 95% confidence), assuming a Gaussian distribution for 15 N CSA values. This number is relatively small, given the ∼30 ppm range of variation in the isotropic chemical shifts, and could be a result of the limited experimental precision in the CSA data, as the experimental uncertainties (±13 ppm) in the individual 15 N CSA values in that paper are noticeably bigger than the reported variability.…”

Section: Introductionsupporting

confidence: 66%

“…Measurements in ubiquitin 5,6 revealed a range of site-specific backbone 15 N CSA values, from approximately -120 to -220 ppm, with a mean of -157 ppm and a standard deviation (not the site-to-site variability) of 19 ppm. This range includes data for both conformationally well-defined amides and those located in the flexible regions.…”

Section: Introductionmentioning

confidence: 96%

“…However, applications of these techniques to uniformly labeled proteins are still in development. Recent solution NMR approaches based on orientation-dependent changes in 15 N resonances in weakly aligned protein solutions 22-24 are very promising, although the accuracy and precision of these measurements do not yet allow site-specific determination of the CST values.…”

Section: Introductionmentioning

confidence: 99%

“…It has been demonstrated [4][5][6]8,25,26 that the anisotropy (CSA), of the 15 N CST can be directly obtained from 15 N relaxation measurements in proteins in solution. Measurements in ubiquitin 5,6 revealed a range of site-specific backbone 15 N CSA values, from approximately -120 to -220 ppm, with a mean of -157 ppm and a standard deviation (not the site-to-site variability) of 19 ppm.…”

Section: Introductionmentioning

confidence: 99%

“…While the existence of some site-specific variability in 15 N CSA is now established, it still remains to be understood whether the differences between the reported data reflect some protein specificity of the CSA distribution or differences in the experimental approaches and/ or in data analyses. Measurements in other proteins and with higher experimental precision are therefore required in order to address this issue.…”

Section: Introductionmentioning

confidence: 99%

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Variability of the ¹⁵N Chemical Shielding Tensors in the B3 Domain of Protein G from ¹⁵N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters

2006

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We applied a combination of 15 N relaxation and CSA/dipolar cross-correlation measurements at five magnetic fields (9.4, 11.7, 14.1, 16.4, and 18.8 Tesla) to determine the 15 N chemical shielding tensors for backbone amides in protein G in solution. The data were analyzed using various modelindependent approaches and those based on Lipari-Szabo approximation, all of them yielding similar results. The results indicate a range of site-specific values of the anisotropy (CSA) and orientation of the 15 N chemical shielding tensor, similar to those in ubiquitin. Assuming a Gaussian distribution of the 15 N CSA values, the mean anisotropy is -173.9 to -177.2 ppm (for 1.02-Å NH-bond length) and the site-so-site CSA variability is ±17.6 to ±21.4 ppm, depending on the method used. This CSA variability is significantly larger than derived previously for ribonuclease H or recently, using "metaanalysis" for ubiquitin. Standard interpretation of 15 N relaxation studies of backbone dynamics in proteins involves an a priori assumption of a uniform 15 N CSA. We show that this assumption leads to a significant discrepancy between the order parameters obtained at different fields. Using the sitespecific CSAs obtained from our study removes this discrepancy and allows simultaneous fit of relaxation data at all five fields to Lipari-Szabo spectral densities. These findings emphasize the necessity of taking into account the variability of 15 N CSA for accurate analysis of protein dynamics from 15 N relaxation measurements.

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Section: Introductionsupporting

confidence: 66%

Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Variability of the ¹⁵N Chemical Shielding Tensors in the B3 Domain of Protein G from ¹⁵N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters

2006

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Structure and dynamics of the M13 coat signal sequence in membranes by multidimensional high-resolution and solid-state NMR spectroscopy

Bechinger

1997

Proteins

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The polypeptide corresponding to the signal sequence of the M13 coat protein and the five N-terminal residues of the mature protein was prepared by solid-phase peptide synthesis with a 15N isotopic label at the alanine-12 position. Multidimensional solution NMR spectroscopy and molecular modeling calculations indicate that this polypeptide assumes helical conformations between residues 5 and 20, in the presence of sodium dodecylsulfate micelles. This is in good agreement with circular dichroism spectroscopic measurement, which shows an alpha-helix content of approximately 42%. The alpha-helix comprises an uninterrupted hydrophobic stretch of < or = 12 amino acids, which is generally believed to be too short for a stable transmembrane alignment in a biological bilayer. The monoexponential proton-deuterium exchange kinetics of this hydrophobic helical region is characterized by half-lives of 15-75 minutes (pH 4.2, 323 K). When the polypeptide is reconstituted into phospholipid bilayers, the broad anisotropy of the proton-decoupled 15N solid-state NMR spectroscopy indicates that the hydrophobic helix is immobilized close to the lipid bilayer surface at the time scale of 15N solid-state NMR spectroscopy (10(-4) seconds). By contrast, short correlation times, immediate hydrogen-deuterium exchange as well as nuclear Overhauser effect crosspeak analysis suggest that the N and C termini of this polypeptide exhibit a mobile random coil structure. The implications of these structural findings for possible mechanisms of membrane insertion and translocation as well as for membrane protein structure prediction algorithms are discussed.

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Protein structure in anisotropic environments: Development of orientational constraints

Cross

Quine

2000

Concepts Magn. Reson.

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15N chemical shift and 15N-13C dipolar tensors for the peptide bond in [1-13C]glycyl[15N]glycine hydrochloride monohydrate

Cited by 75 publications

References 11 publications

Variability of the ¹⁵N Chemical Shielding Tensors in the B3 Domain of Protein G from ¹⁵N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters

Variability of the ¹⁵N Chemical Shielding Tensors in the B3 Domain of Protein G from ¹⁵N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters

Structure and dynamics of the M13 coat signal sequence in membranes by multidimensional high-resolution and solid-state NMR spectroscopy

Protein structure in anisotropic environments: Development of orientational constraints

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15N chemical shift and 15N-13C dipolar tensors for the peptide bond in [1-13C]glycyl[15N]glycine hydrochloride monohydrate

Cited by 75 publications

References 11 publications

Variability of the 15N Chemical Shielding Tensors in the B3 Domain of Protein G from 15N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters

Variability of the 15N Chemical Shielding Tensors in the B3 Domain of Protein G from 15N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters

Structure and dynamics of the M13 coat signal sequence in membranes by multidimensional high-resolution and solid-state NMR spectroscopy

Protein structure in anisotropic environments: Development of orientational constraints

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Variability of the ¹⁵N Chemical Shielding Tensors in the B3 Domain of Protein G from ¹⁵N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters

Variability of the ¹⁵N Chemical Shielding Tensors in the B3 Domain of Protein G from ¹⁵N Relaxation Measurements at Several Fields. Implications for Backbone Order Parameters