TY - JOUR

T1 - Peptide discs as precursors of biologically relevant supported lipid bilayers

AU - Luchini, Alessandra

AU - Sebastiani, Federica

AU - Tidemand, Frederik Grønbæk

AU - Batchu, Krishna Chaithanya

AU - Campana, Mario

AU - Fragneto, Giovanna

AU - Cárdenas, Marité

AU - Arleth, Lise

N1 - Funding Information: The work was supported by grants from Novo Nordisk foundation Interdisciplinary Synergy program, the Lundbeck foundation “BRAINSTRUC” project and Danscatt for travel support. The authors also thank the ISIS neutron source (10.5286/ISIS.E.RB1910248, 10.5286/ISIS.E.RB1920320) and the Institut Laue Langevin (ILL) (10.5291/ILL-DATA.8-02-874) for the allocation of beamtime and Dr. Samantha Micciulla for technical support during the NR experiments at ILL. The authors thank the Deuteration lab and Valerie Laux for the preparation of the hydrogenous and natural yeast cells and the Partnership for Soft Condensed Matter (PSCM) for the lab support during the neutron reflectometry experiments. Lipid production at the ILL was funded by the SINE2020 European Union project within the European Commission's Horizon 2020 Research and Innovation programme under the grant agreements N°64500. AL also thanks Prof. Paolo Luchini for the useful discussions during the development of the code for NR data analysis. Funding Information: The work was supported by grants from Novo Nordisk foundation Interdisciplinary Synergy program, the Lundbeck foundation “BRAINSTRUC” project and Danscatt for travel support. The authors also thank the ISIS neutron source (10.5286/ISIS.E.RB1910248, 10.5286/ISIS.E.RB1920320) and the Institut Laue Langevin (ILL) (10.5291/ILL-DATA.8-02-874) for the allocation of beamtime and Dr. Samantha Micciulla for technical support during the NR experiments at ILL. The authors thank the Deuteration lab and Valerie Laux for the preparation of the hydrogenous and natural yeast cells and the Partnership for Soft Condensed Matter (PSCM) for the lab support during the neutron reflectometry experiments. Lipid production at the ILL was funded by the SINE2020 European Union project within the European Commission's Horizon 2020 Research and Innovation programme under the grant agreements N°64500. AL also thanks Prof. Paolo Luchini for the useful discussions during the development of the code for NR data analysis. Publisher Copyright: © 2020 Elsevier Inc.

PY - 2021/3

Y1 - 2021/3

N2 - Supported lipid bilayers (SLBs) are commonly used to investigate the structure and dynamics of biological membranes. Vesicle fusion is a widely exploited method to produce SLBs. However, this process becomes less favoured when the vesicles contain complex lipid mixtures, e.g. natural lipid extracts. In these cases, it is often necessary to change experimental parameters, such as temperature, to unphysiological values to trigger the SLB formation. This may induce lipid degradation and is also not compatible with including membrane proteins or other biomolecules into the bilayers. Here, we show that the peptide discs, ~10 nm discoidal lipid bilayers stabilized in solution by a self-assembled 18A peptide belt, can be used as precursors for SLBs. The characterizations by means of neutron reflectometry and attenuated total reflectance-FTIR spectroscopy show that SLBs were successfully formed both from synthetic lipid mixtures (surface coverage 90–95%) and from natural lipid mixtures (surface coverage ~85%). Traces of 18A peptide (below 0.02 M ratio) left at the support surface after the bilayer formation do not affect the SLB structure. Altogether, we demonstrate that peptide disc formation of SLBs is much faster than the SLB formation by vesicle fusion and without the need of altering any experimental variable from physiologically relevant values.

AB - Supported lipid bilayers (SLBs) are commonly used to investigate the structure and dynamics of biological membranes. Vesicle fusion is a widely exploited method to produce SLBs. However, this process becomes less favoured when the vesicles contain complex lipid mixtures, e.g. natural lipid extracts. In these cases, it is often necessary to change experimental parameters, such as temperature, to unphysiological values to trigger the SLB formation. This may induce lipid degradation and is also not compatible with including membrane proteins or other biomolecules into the bilayers. Here, we show that the peptide discs, ~10 nm discoidal lipid bilayers stabilized in solution by a self-assembled 18A peptide belt, can be used as precursors for SLBs. The characterizations by means of neutron reflectometry and attenuated total reflectance-FTIR spectroscopy show that SLBs were successfully formed both from synthetic lipid mixtures (surface coverage 90–95%) and from natural lipid mixtures (surface coverage ~85%). Traces of 18A peptide (below 0.02 M ratio) left at the support surface after the bilayer formation do not affect the SLB structure. Altogether, we demonstrate that peptide disc formation of SLBs is much faster than the SLB formation by vesicle fusion and without the need of altering any experimental variable from physiologically relevant values.

KW - ATR-FTIR

KW - Neutron reflectometry

KW - Peptide discs

KW - Supported lipid bilayers

U2 - 10.1016/j.jcis.2020.11.086

DO - 10.1016/j.jcis.2020.11.086

M3 - Journal article

C2 - 33307306

VL - 585

SP - 376

EP - 385

JO - Journal of Colloid and Interface Science

JF - Journal of Colloid and Interface Science

SN - 0021-9797

ER -