Characterizing the structure of lipodisq nanoparticles for membrane protein spectroscopic studies.
Membrane protein spectroscopic studies are challenging due to the difficulty introduced in preparing homogenous and functional hydrophobic proteins incorporated into a lipid bilayer system. Traditional membrane mimics such as micelles or liposomes have proved to be powerful in solubilizing membrane proteins for biophysical studies, however, several drawbacks have limited their applications. Recently, a nanosized complex termed lipodisq nanoparticles was utilized as an alternative membrane mimic to overcome these caveats by providing a homogeneous lipid bilayer environment. Despite all the benefits that lipodisq nanoparticles could provide to enhance the biophysical studies of membrane proteins, structural characterization in different lipid compositions that closely mimic the native membrane environment is still lacking. In this study, the formation of lipodisq nanoparticles using different weight ratios of POPC/POPG lipids to SMA polymers was characterized via solid-state nuclear magnetic resonance (SSNMR) spectroscopy and dynamic light scattering (DLS). A critical weight ratio of (1/1.25) for the complete solubilization of POPC/POPG vesicles has been observed and POPC/POPG vesicles turned clear instantaneously upon the addition of the SMA polymer. The size of lipodisq nanoparticles formed from POPC/POPG lipids at this weight ratio of (1/1.25) was found to be about 30 nm in radius. We also showed that upon the complete solubilization of POPC/POPG vesicles by SMA polymers, the average size of the lipodisq nanoparticles is weight ratio dependent, when more SMA polymers were introduced, smaller lipodisq nanoparticles were obtained. The results of this study will be helpful for a variety of biophysical experiments when specific size of lipid disc is required. Further, this study will provide a proper path for researchers working on membrane proteins to obtain pertinent structure and dynamic information in a physiologically relevant membrane mimetic environment.
Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA.
Rongfu
R
Zhang
Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA.
Indra D
ID
Sahu
Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA.
Lishan
L
Liu
Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA.
Anna
A
Osatuke
Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA.
Raven G
RG
Comer
Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA.
Carole
C
Dabney-Smith
Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA. Electronic address: gary.lorigan@miamioh.edu.
Gary A
GA
Lorigan
D008051:Lipid Bilayers / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008298:Maleates / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry
D008051:Lipid Bilayers; D008298:Maleates; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D011137:Polystyrenes; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; C024075:styrofoam; C030272:maleic acid; C028694:1-palmitoyl-2-oleoylphosphatidylcholine
3:1 SMA polymer; POPC/POPG vesicle; SSNMR spectroscopy; dynamic light scattering; lipodisq nanoparticle