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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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DOI

10.1007/978-3-540-75999-7_8

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1573-5001

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Autoinducing media for uniform isotope labeling o

Authors: Nicolas Guthertz Julia Klopp Aurélie Winterhalter César Fernández Alvar D Gossert
Publish Date: 2015/04/19
Volume: 62, Issue: 2, Pages: 169-177
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Abstract

Autoinducing media for protein expression offer many advantages like robust reproducibility high yields of soluble protein and much reduced workload Here an autoinducing medium for uniform isotope labelling of proteins with 15N 13C and/or 2H in E coli is presented So far autoinducing media have not found widespread application in the NMR field because of the prohibitively high cost of labeled lactose which is an essential ingredient of such media Here we propose using lactose that is only selectively labeled on the glucose moiety It can be synthesized from inexpensive and readily available substrates labeled glucose and unlabeled activated galactose With this approach uniformly isotope labeled proteins were expressed in unattended autoinducing cultures with incorporation of 13C 15N of 966  and 2H 15N of 988  With the present protocol the NMR community could profit from the many advantages that autoinducing media offer

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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. Improved NMR experiments with 13 C-isotropic mixing for assignment of aromatic and aliphatic side chains in labeled proteins
  15. Out-and-back 13 C– 13 C scalar transfers in protein resonance assignment by proton-detected solid-state NMR under ultra-fast MAS
  16. Perspectives for sensitivity enhancement in proton-detected solid-state NMR of highly deuterated proteins by preserving water magnetization
  17. A probe to monitor performance of 15 N longitudinal relaxation experiments for proteins in solution
  18. Optimization of NMR spectroscopy of encapsulated proteins dissolved in low viscosity fluids
  19. Use of quantitative 1 H NMR chemical shift changes for ligand docking into barnase
  20. Resolution enhancement by homonuclear J-decoupling: application to three-dimensional solid-state magic angle spinning NMR spectroscopy
  21. Complexity of aromatic ring-flip motions in proteins: Y97 ring dynamics in cytochrome c observed by cross-relaxation suppressed exchange NMR spectroscopy
  22. Hexagonal ice in pure water and biological NMR samples
  23. Erratum to: 13 C α CEST experiment on uniformly 13 C-labeled proteins
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