Subnanometer Structure of an Asymmetric Model Membrane: Interleaflet Coupling Influences Domain Properties
Abstract
Cell membranes possess a complex three-dimensional architecture, including nonrandom lipid lateral organization within the plane of a bilayer leaflet, and compositional asymmetry between the two leaflets. As a result, delineating the membrane structure–function relationship has been a highly challenging task. Even in simplified model systems, the interactions between bilayer leaflets are poorly understood, due in part to the difficulty of preparing asymmetric model membranes that are free from the effects of residual organic solvent or osmotic stress. To address these problems, we have modified a technique for preparing asymmetric large unilamellar vesicles (aLUVs) via cyclodextrin-mediated lipid exchange in order to produce tensionless, solvent-free aLUVs suitable for a range of biophysical studies. Leaflet composition and structure were characterized using isotopic labeling strategies, which allowed us to avoid the use of bulky labels. NMR and gas chromatography provided precise quantification of the extent of lipid exchange and bilayer asymmetry, while small-angle neutron scattering (SANS) was used to resolve bilayer structural features with subnanometer resolution. Isotopically asymmetric POPC vesicles were found to have the same bilayer thickness and area per lipid as symmetric POPC vesicles, demonstrating that the modified exchange protocol preserves native bilayer structure. Partial exchange of DPPC into the outer leafletmore »
- Authors:
-
- Institute of Molecular Biosciences, Biophysics Division, NAWI Graz, University of Graz, Graz 8010, Austria, BioTechMed-Graz, Graz 8010, Austria
- Tri-Institutional PhD Program in Computational Biology and Medicine, Weill Cornell Medical College, New York, New York 10065, United States, Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853, United States
- Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853, United States
- Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York 11794, United States
- Publication Date:
- Research Org.:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source (SNS)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1257005
- Alternate Identifier(s):
- OSTI ID: 1279447
- Grant/Contract Number:
- AC05-00OR22725; FG02-08ER46528
- Resource Type:
- Published Article
- Journal Name:
- Langmuir
- Additional Journal Information:
- Journal Name: Langmuir Journal Volume: 32 Journal Issue: 20; Journal ID: ISSN 0743-7463
- Publisher:
- American Chemical Society
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Heberle, Frederick A., Marquardt, Drew, Doktorova, Milka, Geier, Barbara, Standaert, Robert F., Heftberger, Peter, Kollmitzer, Benjamin, Nickels, Jonathan D., Dick, Robert A., Feigenson, Gerald W., Katsaras, John, London, Erwin, and Pabst, Georg. Subnanometer Structure of an Asymmetric Model Membrane: Interleaflet Coupling Influences Domain Properties. United States: N. p., 2016.
Web. doi:10.1021/acs.langmuir.5b04562.
Heberle, Frederick A., Marquardt, Drew, Doktorova, Milka, Geier, Barbara, Standaert, Robert F., Heftberger, Peter, Kollmitzer, Benjamin, Nickels, Jonathan D., Dick, Robert A., Feigenson, Gerald W., Katsaras, John, London, Erwin, & Pabst, Georg. Subnanometer Structure of an Asymmetric Model Membrane: Interleaflet Coupling Influences Domain Properties. United States. https://doi.org/10.1021/acs.langmuir.5b04562
Heberle, Frederick A., Marquardt, Drew, Doktorova, Milka, Geier, Barbara, Standaert, Robert F., Heftberger, Peter, Kollmitzer, Benjamin, Nickels, Jonathan D., Dick, Robert A., Feigenson, Gerald W., Katsaras, John, London, Erwin, and Pabst, Georg. Mon .
"Subnanometer Structure of an Asymmetric Model Membrane: Interleaflet Coupling Influences Domain Properties". United States. https://doi.org/10.1021/acs.langmuir.5b04562.
@article{osti_1257005,
title = {Subnanometer Structure of an Asymmetric Model Membrane: Interleaflet Coupling Influences Domain Properties},
author = {Heberle, Frederick A. and Marquardt, Drew and Doktorova, Milka and Geier, Barbara and Standaert, Robert F. and Heftberger, Peter and Kollmitzer, Benjamin and Nickels, Jonathan D. and Dick, Robert A. and Feigenson, Gerald W. and Katsaras, John and London, Erwin and Pabst, Georg},
abstractNote = {Cell membranes possess a complex three-dimensional architecture, including nonrandom lipid lateral organization within the plane of a bilayer leaflet, and compositional asymmetry between the two leaflets. As a result, delineating the membrane structure–function relationship has been a highly challenging task. Even in simplified model systems, the interactions between bilayer leaflets are poorly understood, due in part to the difficulty of preparing asymmetric model membranes that are free from the effects of residual organic solvent or osmotic stress. To address these problems, we have modified a technique for preparing asymmetric large unilamellar vesicles (aLUVs) via cyclodextrin-mediated lipid exchange in order to produce tensionless, solvent-free aLUVs suitable for a range of biophysical studies. Leaflet composition and structure were characterized using isotopic labeling strategies, which allowed us to avoid the use of bulky labels. NMR and gas chromatography provided precise quantification of the extent of lipid exchange and bilayer asymmetry, while small-angle neutron scattering (SANS) was used to resolve bilayer structural features with subnanometer resolution. Isotopically asymmetric POPC vesicles were found to have the same bilayer thickness and area per lipid as symmetric POPC vesicles, demonstrating that the modified exchange protocol preserves native bilayer structure. Partial exchange of DPPC into the outer leaflet of POPC vesicles produced chemically asymmetric vesicles with a gel/fluid phase-separated outer leaflet and a uniform, POPC-rich inner leaflet. SANS was able to separately resolve the thicknesses and areas per lipid of coexisting domains, revealing reduced lipid packing density of the outer leaflet DPPC-rich phase compared to typical gel phases. Lastly, our finding that a disordered inner leaflet can partially fluidize ordered outer leaflet domains indicates some degree of interleaflet coupling, and invites speculation on a role for bilayer asymmetry in modulating membrane lateral organization.},
doi = {10.1021/acs.langmuir.5b04562},
journal = {Langmuir},
number = 20,
volume = 32,
place = {United States},
year = {Mon May 16 00:00:00 EDT 2016},
month = {Mon May 16 00:00:00 EDT 2016}
}
https://doi.org/10.1021/acs.langmuir.5b04562
Web of Science
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