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Title: 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 » of membrane peptide binding, conformation, and insertion on lipid asymmetry.« less

Authors:
 [1];  [1];  [1];  [2]; ORCiD logo [3];  [4];  [4];  [4]; ORCiD logo [5]; ORCiD logo [6]; ORCiD logo [7]; ORCiD logo [1]
+ Show Author Affiliations
  1. Univ. of Windsor, Windsor, ON (Canada)
  2. Univ. of Texas Health Science Center at Houston, Houston, TX (United States)
  3. Univ. of Texas Health Science Center at Houston, Houston, TX (United States); Univ. of Tennessee, Knoxville, TN (United States)
  4. Univ. of Tennessee, Knoxville, TN (United States)
  5. Univ. of Tennessee, Knoxville, TN (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  6. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  7. 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.
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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
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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.
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@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}
}
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Journal Article:
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Publisher's Version of Record
https://doi.org/10.1021/acs.langmuir.9b01625
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Figures / Tables:

Figure 1 Figure 1: (a) Cartoon of an asymmetric bilayer. Dark shaded components indicates deuterated moieties. (b) Predicted neutron scattering curves of asymmetric (dH-POPCinn/dC-DMPCout) and symmetric vesicles calculated using a six-slab bilayer model. (c) I(q) scattering data from three different samples: (1) compositionally asymmetric LUVs with their inner leaflets composed mainly ofmore » dHPOPC and outer leafet of dC-DMPC (pink circles); (2) aLUVs with a 1/40 P/L ratio of gA (purple triangles); and (3) uniformly mixed LUVs with same dH-POPC and dC-DMPC ratios (navy squares). Scattering curves are overlaid to highlight similarities and differences.« less
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