Journal of Biomolecular NMR

, Volume 61, Issue 3–4, pp 361–367 | Cite as

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


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.


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



This work was supported by grants from the National Institutes of Health and National Science Foundation. Professor McDermott is a member of the New York Structural Biology Center. The Center is a STAR center supported by the New York State Office of Science, Technology, and Academic Research. NMR resources were supported by NIH P41 GM66354. This work was supported by NSF MCB 0749381, NIH R01 GM 88724 to A. E. M., NIH NRSA F32 087908 to B. J. W. and by NIH/NIGMS P41GM103521 and NIH/NIBIB R01EB003150 to J.H.F.

Supplementary material

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Supplementary material 1 (DOCX 734 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Benjamin J. Wylie
    • 1
  • Boris G. Dzikovski
    • 2
  • Shane Pawsey
    • 3
  • Marc Caporini
    • 3
  • Melanie Rosay
    • 3
  • Jack H. Freed
    • 2
  • Ann E. McDermott
    • 1
  1. 1.Department of ChemistryColumbia UniversityNew YorkUSA
  2. 2.National Biomedical Center for Advanced ESR Technology, Department of Chemistry and Chemical BiologyCornell UniversityIthacaUSA
  3. 3.Bruker BioSpin CorporationBillericaUSA

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