1 H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers

Marquardt, Drew; Heberle, Frederick A.; Miti, Tatiana; Eicher, Barbara; London, Erwin; Katsaras, John; Pabst, Georg

doi:10.1021/acs.langmuir.6b04485

Title: ¹ H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers

Abstract

We measured the transbilayer diffusion of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in large unilamellar vesicles, in both the gel (L_β') and fluid (L_α) phases. The choline resonance of headgroup-protiated DPPC exchanged into the outer leaflet of headgroup-deuterated DPPC-d13 vesicles was monitored using ¹H NMR spectroscopy, coupled with the addition of a paramagnetic shift reagent. This allowed us to distinguish between the inner and outer bilayer leaflet of DPPC, to determine the flip-flop rate as a function of temperature. Flip-flop of fluid-phase DPPC exhibited Arrhenius kinetics, from which we determined an activation energy of 122 kJ mol^–1. In gel-phase DPPC vesicles, flip-flop was not observed over the course of 250 h. Our findings are in contrast to previous studies of solid-supported bilayers, where the reported DPPC translocation rates are at least several orders of magnitude faster than those in vesicles at corresponding temperatures. We reconcile these differences by proposing a defect-mediated acceleration of lipid translocation in supported bilayers, where long-lived, submicron-sized holes resulting from incomplete surface coverage are the sites of rapid transbilayer movement.

Authors:

; Heberle, Frederick A.; Miti, Tatiana; Eicher, Barbara; London, Erwin; Katsaras, John;

Publication Date:: Fri Feb 03 00:00:00 EST 2017

Research Org.:: Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)

Sponsoring Org.:: Austrian Science Fund (FWF); National Science Foundation (NSF); USDOE Office of Science (SC)

OSTI Identifier:: 1342412

Alternate Identifier(s):: OSTI ID: 1352553; OSTI ID: 1362240

Grant/Contract Number:: 7394; AC05-00OR22725

Resource Type:: Published Article

Journal Name:: Langmuir

Additional Journal Information:: Journal Name: Langmuir Journal Volume: 33 Journal Issue: 15; Journal ID: ISSN 0743-7463

Publisher:: American Chemical Society

Country of Publication:: United States

Language:: English

Subject:: 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES

Citation Formats


                    Marquardt, Drew, Heberle, Frederick A., Miti, Tatiana, Eicher, Barbara, London, Erwin, Katsaras, John, and Pabst, Georg. 1 H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers.  United States: N. p., 2017. 
Web.  doi:10.1021/acs.langmuir.6b04485.

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                    Marquardt, Drew, Heberle, Frederick A., Miti, Tatiana, Eicher, Barbara, London, Erwin, Katsaras, John, & Pabst, Georg. 1 H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers.  United States.  https://doi.org/10.1021/acs.langmuir.6b04485

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                    Marquardt, Drew, Heberle, Frederick A., Miti, Tatiana, Eicher, Barbara, London, Erwin, Katsaras, John, and Pabst, Georg. Fri .  
"1 H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers".  United States.  https://doi.org/10.1021/acs.langmuir.6b04485.

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@article{osti_1342412,

  title        = {1 H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers},

  author       = {Marquardt, Drew and Heberle, Frederick A. and Miti, Tatiana and Eicher, Barbara and London, Erwin and Katsaras, John and Pabst, Georg},

  abstractNote = {We measured the transbilayer diffusion of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in large unilamellar vesicles, in both the gel (Lβ') and fluid (Lα) phases. The choline resonance of headgroup-protiated DPPC exchanged into the outer leaflet of headgroup-deuterated DPPC-d13 vesicles was monitored using 1H NMR spectroscopy, coupled with the addition of a paramagnetic shift reagent. This allowed us to distinguish between the inner and outer bilayer leaflet of DPPC, to determine the flip-flop rate as a function of temperature. Flip-flop of fluid-phase DPPC exhibited Arrhenius kinetics, from which we determined an activation energy of 122 kJ mol–1. In gel-phase DPPC vesicles, flip-flop was not observed over the course of 250 h. Our findings are in contrast to previous studies of solid-supported bilayers, where the reported DPPC translocation rates are at least several orders of magnitude faster than those in vesicles at corresponding temperatures. We reconcile these differences by proposing a defect-mediated acceleration of lipid translocation in supported bilayers, where long-lived, submicron-sized holes resulting from incomplete surface coverage are the sites of rapid transbilayer movement.},

  doi          = {10.1021/acs.langmuir.6b04485},

  journal      = {Langmuir},

  number       = 15,

  volume       = 33,

  place        = {United States},

  year         = {Fri Feb 03 00:00:00 EST 2017},

  month        = {Fri Feb 03 00:00:00 EST 2017}

}

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Works referencing / citing this record:

Complex biomembrane mimetics on the sub-nanometer scale
journal, July 2017

Heberle, Frederick A.; Pabst, Georg
Biophysical Reviews, Vol. 9, Issue 4
DOI: 10.1007/s12551-017-0275-5

Preparation of asymmetric phospholipid vesicles for use as cell membrane models
journal, September 2018

Doktorova, Milka; Heberle, Frederick A.; Eicher, Barbara
Nature Protocols, Vol. 13, Issue 9
DOI: 10.1038/s41596-018-0033-6

Transverse lipid organization dictates bending fluctuations in model plasma membranes
journal, January 2020

Rickeard, Brett W.; Nguyen, Michael H. L.; DiPasquale, Mitchell
Nanoscale, Vol. 12, Issue 3
DOI: 10.1039/c9nr07977g

Lipid diffusion in the distal and proximal leaflets of supported lipid bilayer membranes studied by single particle tracking
journal, March 2018

Schoch, Rafael L.; Barel, Itay; Brown, Frank L. H.
The Journal of Chemical Physics, Vol. 148, Issue 12
DOI: 10.1063/1.5010341

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Title: 1 H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers

Abstract

Citation Formats

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Preparation of asymmetric phospholipid vesicles for use as cell membrane models journal, September 2018

Transverse lipid organization dictates bending fluctuations in model plasma membranes journal, January 2020

Lipid diffusion in the distal and proximal leaflets of supported lipid bilayer membranes studied by single particle tracking journal, March 2018

Title: ¹ H NMR Shows Slow Phospholipid Flip-Flop in Gel and Fluid Bilayers

Complex biomembrane mimetics on the sub-nanometer scale
journal, July 2017

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journal, September 2018

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journal, January 2020

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journal, March 2018