Journal of Biomolecular NMR

, Volume 18, Issue 4, pp 303–309 | Cite as

Solution NMR of proteins within polyacrylamide gels: Diffusional properties and residual alignment by mechanical stress or embedding of oriented purple membranes

  • Hans-Jürgen Sass
  • Giovanna Musco
  • Stephen J. Stahl
  • Paul T. Wingfield
  • Stephan Grzesiek

Abstract

The diffusive properties of biomacromolecules within the aqueous phase of polyacrylamide gels are described. High quality NMR spectra can be obtained under such conditions. As compared to water, a fivefold reduction in the translational diffusion constant, but only a 1.6-fold decrease (1.4-fold increase) in amide-15N T2 (T1) are observed for human ubiquitin within a 10% acrylamide gel. Weak alignment of the solute macromolecules can be achieved within such gels by vertical or radial compression or by the embedding of magnetically oriented purple membrane fragments. The methods are applied to derive residual dipolar couplings for human HIV-1 Nef and ubiquitin.

dipolar couplings HIV-1 Nef ubiquitin 

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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Hans-Jürgen Sass
    • 1
  • Giovanna Musco
    • 1
  • Stephen J. Stahl
    • 2
  • Paul T. Wingfield
    • 2
  • Stephan Grzesiek
    • 1
  1. 1.Department of Structural Biology, BiozentrumUniversity of BaselBaselSwitzerland
  2. 2.National Institute of Arthritis and Musculoskeletal and Skin DiseasesNational Institutes of HealthBethesdaU.S.A.

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