title: Dynamic nuclear polarization of membrane proteins: covalently bound spin-labels at protein-protein interfaces.
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.
authors.affiliation: Department of Chemistry, Columbia University, New York, NY, 10027, USA.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Wylie
authors.forename: Boris G
authors.initials: BG
authors.lastname: Dzikovski
authors.forename: Shane
authors.initials: S
authors.lastname: Pawsey
authors.forename: Marc
authors.initials: M
authors.lastname: Caporini
authors.forename: Melanie
authors.initials: M
authors.lastname: Rosay
authors.forename: Jack H
authors.initials: JH
authors.lastname: Freed
authors.forename: Ann E
authors.initials: AE
authors.lastname: McDermott
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D006096:Gramicidin / Q000737:chemistry / Q000378:metabolism*; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry; D054730:Protein Interaction Domains and Motifs; D013113:Spin Labels
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D008051:Lipid Bilayers; D008565:Membrane Proteins; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D013113:Spin Labels; D006096:Gramicidin; C038089:1,2-distearoylphosphatidylethanolamine; C042511:1,2-dioleoylphosphatidylserine
references.citation: J Biomol NMR. 1996 Jul;8(1):1-14
references.pmid: 8810522
references.citation: Appl Magn Reson. 2008 Aug;34(3-4):237-263
references.pmid: 19194532
references.citation: Science. 1997 May 9;276(5314):930-2
references.pmid: 9139651
references.citation: Biophys J. 2004 Nov;87(5):3504-17
references.pmid: 15326023
references.citation: Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8239-43
references.pmid: 7667275
references.citation: J Phys Chem B. 2011 Jan 13;115(1):176-85
references.pmid: 21142163
references.citation: J Am Chem Soc. 2004 Sep 8;126(35):10844-5
references.pmid: 15339160
references.citation: J Am Chem Soc. 2006 Sep 6;128(35):11385-90
references.pmid: 16939261
references.citation: Biophys J. 1998 Nov;75(5):2478-88
references.pmid: 9788943
references.citation: Annu Rev Biophys. 2009;38:385-403
references.pmid: 19245337
references.citation: Solid State Nucl Magn Reson. 2012 May-Jun;43-44:1-13
references.pmid: 22464402
references.citation: Chemphyschem. 2011 Nov 18;12(16):2929-32
references.pmid: 22028312
references.citation: Chem Rev. 2004 Aug;104(8):3587-606
references.pmid: 15303829
references.citation: J Chem Phys. 2008 Feb 7;128(5):052302
references.pmid: 18266419
references.citation: J Am Chem Soc. 2003 Nov 12;125(45):13626-7
references.pmid: 14599177
references.citation: Angew Chem Int Ed Engl. 2009;48(27):4996-5000
references.pmid: 19492374
references.citation: J Am Chem Soc. 2011 Dec 7;133(48):19266-9
references.pmid: 22039931
references.citation: J Am Chem Soc. 2007 Jun 20;129(24):7502-3
references.pmid: 17530852
references.citation: Proc Natl Acad Sci U S A. 2009 Jun 9;106(23):9244-9
references.pmid: 19474298
references.citation: Annu Rev Phys Chem. 2012;63:1-24
references.pmid: 22136620
references.citation: Biochim Biophys Acta. 2011 Aug;1808(8):1957-74
references.pmid: 21477581
references.citation: J Magn Reson. 2007 Dec;189(2):251-79
references.pmid: 17942352
references.citation: Anal Biochem. 1978 Jun 1;86(2):512-8
references.pmid: 77645