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Title of Journal: J Biomol NMR

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Abbravation: Journal of Biomolecular NMR

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Springer Netherlands

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10.1007/bf01026711

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1573-5001

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Improved NMR experiments with Superscript13/Sup

Authors: Helena Kovacs Alvar Gossert
Publish Date: 2014/01/04
Volume: 58, Issue: 2, Pages: 101-112
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Abstract

Three improved 13Cspinlock experiments for side chain assignments of isotope labelled proteins in liquid state are presented These are based on wide bandwidth spinlock techniques that have become possible with contemporary cryogenic probes The first application the HCaliCaroHTOCSY is an HCCHTOCSY in which all CHn moieties of a protein are detected in a single experiment including the aromatic ones This enables unambiguous assignment of aromatic and aliphatic amino acids in a single highly sensitive experiment In the second application the 13Cdetected CallTOCSY magnetization transfer comprises all carbons—aliphatic aromatic as well as the carbonyl carbons—making the complete carbon assignment possible using one spectrum only Thirdly the frequently used HCCCONH experiment was redesigned by replacing the long Ccarbonyl refocused INEPT transfer step by direct 13C–13CTOCSY magnetization transfer from side chain carbons to the backbone carbonyls The resulting HCCCONH experiment minimizes relaxation losses because it is shorter and represents a more sensitive alternative particularly for larger proteins The performance of the experiments is demonstrated on isotope labeled proteins up to the size of 43 kDaWe would like to thank Dr Ēriks Kupče at Bruker BioSpin UK for his kind help in creating an appropriate adiabatic mixing pulse and Dr Wolfgang Bermel Bruker BioSpin Germany for perfecting the pulse sequences We thank Julia Klopp and Aurélie Winterhalter for production of the MBP sample


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Other Papers In This Journal:

  1. Detection of nanosecond time scale side-chain jumps in a protein dissolved in water/glycerol solvent
  2. Statistical removal of background signals from high-throughput 1 H NMR line-broadening ligand-affinity screens
  3. Peak picking multidimensional NMR spectra with the contour geometry based algorithm CYPICK
  4. HNHC: a triple resonance experiment for correlating the H2, N1(N3) and C2 resonances in adenine nucleobases of 13 C-, 15 N-labeled RNA oligonucleotides
  5. NOESY-WaterControl: a new NOESY sequence for the observation of under-water protein resonances
  6. Insights into furanose solution conformations: beyond the two-state model
  7. Selective 1 H- 13 C NMR spectroscopy of methyl groups in residually protonated samples of large proteins
  8. Fast methionine-based solution structure determination of calcium-calmodulin complexes
  9. A procedure to validate and correct the 13 C chemical shift calibration of RNA datasets
  10. Complete dissociation of the HIV-1 gp41 ectodomain and membrane proximal regions upon phospholipid binding
  11. NMR solution structure of the acylphosphatase from Escherichia coli
  12. A rigid disulfide-linked nitroxide side chain simplifies the quantitative analysis of PRE data
  13. S3EPY: a Sparky extension for determination of small scalar couplings from spin-state-selective excitation NMR experiments
  14. Out-and-back 13 C– 13 C scalar transfers in protein resonance assignment by proton-detected solid-state NMR under ultra-fast MAS
  15. Perspectives for sensitivity enhancement in proton-detected solid-state NMR of highly deuterated proteins by preserving water magnetization
  16. A probe to monitor performance of 15 N longitudinal relaxation experiments for proteins in solution
  17. Optimization of NMR spectroscopy of encapsulated proteins dissolved in low viscosity fluids
  18. Use of quantitative 1 H NMR chemical shift changes for ligand docking into barnase
  19. Resolution enhancement by homonuclear J-decoupling: application to three-dimensional solid-state magic angle spinning NMR spectroscopy
  20. Complexity of aromatic ring-flip motions in proteins: Y97 ring dynamics in cytochrome c observed by cross-relaxation suppressed exchange NMR spectroscopy
  21. Hexagonal ice in pure water and biological NMR samples
  22. Erratum to: 13 C α CEST experiment on uniformly 13 C-labeled proteins
  23. Auto-inducing media for uniform isotope labeling of proteins with 15 N, 13 C and 2 H

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