Paper Search Console

Home Search Page About Contact

Journal Title

Title of Journal: J Biomol NMR

Search In Journal Title:

Abbravation: Journal of Biomolecular NMR

Search In Journal Abbravation:

Publisher

Springer Netherlands

Search In Publisher:

DOI

10.1007/bf01561059

Search In DOI:

ISSN

1573-5001

Search In ISSN:
Search In Title Of Papers:

Optimization of NMR spectroscopy of encapsulated p

Authors: Nathaniel V Nucci Bryan S Marques Sabrina Bédard Jakob Dogan John M Gledhill Veronica R Moorman Ronald W Peterson Kathleen G Valentine Alison L Wand A Joshua Wand
Publish Date: 2011/07/12
Volume: 50, Issue: 4, Pages: 421-
PDF Link

Abstract

Comprehensive application of solution NMR spectroscopy to studies of macromolecules remains fundamentally limited by the molecular rotational correlation time For proteins molecules larger than 30 kDa require complex experimental methods such as TROSY in conjunction with isotopic labeling schemes that are often expensive and generally reduce the potential information available We have developed the reverse micelle encapsulation strategy as an alternative approach Encapsulation of proteins within the protective nanoscale water pool of a reverse micelle dissolved in ultralow viscosity nonpolar solvents overcomes the slow tumbling problem presented by large proteins Here we characterize the contributions from the various components of the proteincontaining reverse micelle system to the rotational correlation time of the encapsulated protein Importantly we demonstrate that the protein encapsulated in the reverse micelle maintains a hydration shell comparable in size to that seen in bulk solution Using moderate pressures encapsulation in ultralow viscosity propane or ethane can be used to magnify this advantage We show that encapsulation in liquid ethane can be used to reduce the tumbling time of the 43 kDa maltose binding protein from ~23 to ~10 ns These conditions enable for example acquisition of TOCSYtype data resolved on the adjacent amide NH for the 43 kDa encapsulated maltose binding protein dissolved in liquid ethane which is typically impossible for proteins of such size without use of extensive deuteration or the TROSY effectSupported by NSF grants MCB0842814 and DMR05200020 and NIH grant GM 085120 and NIH postdoctoral fellowship GM087099 to NVN JD acknowledges financial support from the WennerGren Foundations AJW and RWP declare a financial conflict of interest as Members of Daeadalus Innovations LLC a manufacturer of highpressure and reverse micelle NMR apparatus


Keywords:

References


.
Search In Abstract Of Papers:
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. 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

Search Result: