Dynamic nuclear polarization of membrane proteins: Covalently bound spin-labels at protein-protein interfaces

Benjamin J. Wylie, Boris G. Dzikovski, Shane Pawsey, Marc Caporini, Melanie Rosay, Jack H. Freed, Ann E. McDermott

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

We demonstrate that dynamic nuclear polarization of membrane proteins in lipid bilayers may be achieved using a novel polarizing agent: pairs of spin labels covalently bound to a protein of interest interacting at an intermolecular interaction surface. For gramicidin A, nitroxide tags attached to the N-terminal intermolecular interface region become proximal only when bimolecular channels forms in the membrane. We obtained signal enhancements of sixfold for the dimeric protein. The enhancement effect was comparable to that of a doubly tagged sample of gramicidin C, with intramolecular spin pairs. This approach could be a powerful and selective means for signal enhancement in membrane proteins, and for recognizing intermolecular interfaces.

Original languageEnglish
Pages (from-to)361-367
Number of pages7
JournalJournal of Biomolecular NMR
Volume61
Issue number3-4
DOIs
StatePublished - Apr 1 2015

Keywords

  • DEER spectroscopy
  • DNP
  • Dynamic nuclear polarization
  • Gramicidin
  • Membrane proteins
  • Solid-state NMR

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