Optimized reverse micelle surfactant system for high-resolution NMR spectroscopy of encapsulated proteins and nucleic acids dissolved in low viscosity fluids.
An optimized reverse micelle surfactant system has been developed for solution nuclear magnetic resonance studies of encapsulated proteins and nucleic acids dissolved in low viscosity fluids. Comprising the nonionic 1-decanoyl-rac-glycerol and the zwitterionic lauryldimethylamine-N-oxide (10MAG/LDAO), this mixture is shown to efficiently encapsulate a diverse set of proteins and nucleic acids. Chemical shift analyses of these systems show that high structural fidelity is achieved upon encapsulation. The 10MAG/LDAO surfactant system reduces the molecular reorientation time for encapsulated macromolecules larger than ~20 kDa leading to improved overall NMR performance. The 10MAG/LDAO system can also be used for solution NMR studies of lipid-modified proteins. New and efficient strategies for optimization of encapsulation conditions are described. 10MAG/LDAO performs well in both the low viscosity pentane and ultralow viscosity liquid ethane and therefore will serve as a general surfactant system for initiating solution NMR studies of proteins and nucleic acids.
Johnson Research Foundation and Department of Biochemistry and Biophysics, University of Pennsylvania , Philadelphia, Pennsylvania 19104-6059.
Igor
I
Dodevski
Nathaniel V
NV
Nucci
Kathleen G
KG
Valentine
Gurnimrat K
GK
Sidhu
Evan S
ES
O'Brien
Arthur
A
Pardi
A Joshua
AJ
Wand
D002214:Capsules; D004247:DNA / Q000737:chemistry*; D004123:Dimethylamines / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry*; D008823:Micelles; D008958:Models, Molecular; D008968:Molecular Conformation; D012313:RNA / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*; D014783:Viscosity; D014835:Volatilization
D002214:Capsules; D004123:Dimethylamines; D008565:Membrane Proteins; D008823:Micelles; D013501:Surface-Active Agents; C014518:dodecyldimethylamine oxide; D012313:RNA; D004247:DNA