Peptide-Induced Lipid Flip-Flop in Asymmetric Liposomes Measured by Small Angle Neutron Scattering
Abstract
Despite the prevalence of lipid transbilayer asymmetry in natural plasma membranes, most biomimetic model membranes studied are symmetric. Recent advances have helped to overcome the difficulties in preparing asymmetric liposomes in vitro, allowing for the examination of a larger set of relevant biophysical questions. Here, we investigate the stability of asymmetric bilayers by measuring lipid flip-flop with time-resolved small-angle neutron scattering (SANS). Asymmetric large unilamellar vesicles with inner bilayer leaflets containing predominantly 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and outer leaflets composed mainly of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) displayed slow spontaneous flip-flop at 37 °C (half-time, t1/2 = 140 h). However, inclusion of peptides, namely, gramicidin, alamethicin, melittin, or pHLIP (i.e., pH-low insertion peptide), accelerated lipid flip-flop. For three of these peptides (i.e., pHLIP, alamethicin, and melittin), each of which was added externally to preformed asymmetric vesicles, we observed a completely scrambled bilayer in less than 2 h. Gramicidin, on the other hand, was preincorporated during the formation of the asymmetric liposomes and showed a time resolvable 8-fold increase in the rate of lipid asymmetry loss. These results point to a membrane surface-related (e.g., adsorption/insertion) event as the primary driver of lipid scrambling in the asymmetric model membranes of this study. Here, we discuss the implicationsmore »
- Authors:
-
- Univ. of Windsor, Windsor, ON (Canada)
- Univ. of Texas Health Science Center at Houston, Houston, TX (United States)
- Univ. of Texas Health Science Center at Houston, Houston, TX (United States); Univ. of Tennessee, Knoxville, TN (United States)
- Univ. of Tennessee, Knoxville, TN (United States)
- Univ. of Tennessee, Knoxville, TN (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1561589
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Langmuir
- Additional Journal Information:
- Journal Volume: 35; Journal Issue: 36; Journal ID: ISSN 0743-7463
- Publisher:
- American Chemical Society
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Citation Formats
Nguyen, Michael H. L., DiPasquale, Mitchell, Rickeard, Brett W., Doktorova, Milka, Heberle, Frederick A., Scott, Haden L., Barrera, Francisco N., Taylor, Graham, Collier, Charles P., Stanley, Christopher B., Katsaras, John, and Marquardt, Drew. Peptide-Induced Lipid Flip-Flop in Asymmetric Liposomes Measured by Small Angle Neutron Scattering. United States: N. p., 2019.
Web. doi:10.1021/acs.langmuir.9b01625.
Nguyen, Michael H. L., DiPasquale, Mitchell, Rickeard, Brett W., Doktorova, Milka, Heberle, Frederick A., Scott, Haden L., Barrera, Francisco N., Taylor, Graham, Collier, Charles P., Stanley, Christopher B., Katsaras, John, & Marquardt, Drew. Peptide-Induced Lipid Flip-Flop in Asymmetric Liposomes Measured by Small Angle Neutron Scattering. United States. https://doi.org/10.1021/acs.langmuir.9b01625
Nguyen, Michael H. L., DiPasquale, Mitchell, Rickeard, Brett W., Doktorova, Milka, Heberle, Frederick A., Scott, Haden L., Barrera, Francisco N., Taylor, Graham, Collier, Charles P., Stanley, Christopher B., Katsaras, John, and Marquardt, Drew. Tue .
"Peptide-Induced Lipid Flip-Flop in Asymmetric Liposomes Measured by Small Angle Neutron Scattering". United States. https://doi.org/10.1021/acs.langmuir.9b01625. https://www.osti.gov/servlets/purl/1561589.
@article{osti_1561589,
title = {Peptide-Induced Lipid Flip-Flop in Asymmetric Liposomes Measured by Small Angle Neutron Scattering},
author = {Nguyen, Michael H. L. and DiPasquale, Mitchell and Rickeard, Brett W. and Doktorova, Milka and Heberle, Frederick A. and Scott, Haden L. and Barrera, Francisco N. and Taylor, Graham and Collier, Charles P. and Stanley, Christopher B. and Katsaras, John and Marquardt, Drew},
abstractNote = {Despite the prevalence of lipid transbilayer asymmetry in natural plasma membranes, most biomimetic model membranes studied are symmetric. Recent advances have helped to overcome the difficulties in preparing asymmetric liposomes in vitro, allowing for the examination of a larger set of relevant biophysical questions. Here, we investigate the stability of asymmetric bilayers by measuring lipid flip-flop with time-resolved small-angle neutron scattering (SANS). Asymmetric large unilamellar vesicles with inner bilayer leaflets containing predominantly 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and outer leaflets composed mainly of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) displayed slow spontaneous flip-flop at 37 °C (half-time, t1/2 = 140 h). However, inclusion of peptides, namely, gramicidin, alamethicin, melittin, or pHLIP (i.e., pH-low insertion peptide), accelerated lipid flip-flop. For three of these peptides (i.e., pHLIP, alamethicin, and melittin), each of which was added externally to preformed asymmetric vesicles, we observed a completely scrambled bilayer in less than 2 h. Gramicidin, on the other hand, was preincorporated during the formation of the asymmetric liposomes and showed a time resolvable 8-fold increase in the rate of lipid asymmetry loss. These results point to a membrane surface-related (e.g., adsorption/insertion) event as the primary driver of lipid scrambling in the asymmetric model membranes of this study. Here, we discuss the implications of membrane peptide binding, conformation, and insertion on lipid asymmetry.},
doi = {10.1021/acs.langmuir.9b01625},
journal = {Langmuir},
number = 36,
volume = 35,
place = {United States},
year = {Tue Aug 13 00:00:00 EDT 2019},
month = {Tue Aug 13 00:00:00 EDT 2019}
}
Web of Science
Figures / Tables:
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