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

H. J. Sass, G. Musco, S. J. Stahl, P. T. Wingfield, S. Grzesiek

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)303-309
Number of pages7
JournalJournal of Biomolecular NMR
Volume18
Issue number4
DOIs
Publication statusPublished - 2000

Keywords

  • Dipolar couplings
  • HIV-1 Nef
  • Ubiquitin

ASJC Scopus subject areas

  • Spectroscopy
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

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