gramicidin-a and 1-2-distearoylphosphatidylethanolamine

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.

Topics: Electron Spin Resonance Spectroscopy; Gramicidin; Lipid Bilayers; Membrane Proteins; Nuclear Magnetic Resonance, Biomolecular; Phosphatidylethanolamines; Phosphatidylserines; Protein Interaction Domains and Motifs; Spin Labels