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Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2)

Supported lipid bilayers (SLBs) are commonly used as model systems mimicking biological membranes. Recently, we reported a new method to produce SLBs with incorporated membrane proteins, which is based on the application of peptide discs [Luchini et al., Analytical Chemistry, 2020, 92, 1081–1088]. P...

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Autores principales: Luchini, Alessandra, Tidemand, Frederik Grønbæk, Johansen, Nicolai Tidemand, Sebastiani, Federica, Corucci, Giacomo, Fragneto, Giovanna, Cárdenas, Marité, Arleth, Lise
Formato: Online Artículo Texto
Lenguaje:English
Publicado: RSC 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9595196/
https://www.ncbi.nlm.nih.gov/pubmed/36341300
http://dx.doi.org/10.1039/d2na00384h
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author Luchini, Alessandra
Tidemand, Frederik Grønbæk
Johansen, Nicolai Tidemand
Sebastiani, Federica
Corucci, Giacomo
Fragneto, Giovanna
Cárdenas, Marité
Arleth, Lise
author_facet Luchini, Alessandra
Tidemand, Frederik Grønbæk
Johansen, Nicolai Tidemand
Sebastiani, Federica
Corucci, Giacomo
Fragneto, Giovanna
Cárdenas, Marité
Arleth, Lise
author_sort Luchini, Alessandra
collection PubMed
description Supported lipid bilayers (SLBs) are commonly used as model systems mimicking biological membranes. Recently, we reported a new method to produce SLBs with incorporated membrane proteins, which is based on the application of peptide discs [Luchini et al., Analytical Chemistry, 2020, 92, 1081–1088]. Peptide discs are small discoidal particles composed of a lipid core and an outer belt of self-assembled 18A peptides. SLBs including membrane proteins can be formed by depositing the peptide discs on a solid support and subsequently removing the peptide by buffer rinsing. Here, we introduce a new variant of the 18A peptide, named dark peptide (d18A). d18A exhibits UV absorption at 214 nm, whereas the absorption at 280 nm is negligible. This improves sample preparation as it enables a direct quantification of the membrane protein concentration in the peptide discs by measuring UV absorption at 280 nm. We describe the application of the peptide discs prepared with d18A (dark peptide discs) to produce SLBs with a membrane protein, synaptobrevin 2 (VAMP2). The collected data showed the successful formation of SLBs with high surface coverage and incorporation of VAMP2 in a single orientation with the extramembrane domain exposed towards the bulk solvent. Compared to 18A, we found that d18A was more efficiently removed from the SLB. Our data confirmed the structural organisation of VAMP2 as including both α-helical and β-sheet secondary structure. We further verified the orientation of VAMP2 in the SLBs by characterising the binding of VAMP2 with α-synuclein. These results point at the produced SLBs as relevant membrane models for biophysical studies as well as nanostructured biomaterials.
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spelling pubmed-95951962022-11-04 Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2) Luchini, Alessandra Tidemand, Frederik Grønbæk Johansen, Nicolai Tidemand Sebastiani, Federica Corucci, Giacomo Fragneto, Giovanna Cárdenas, Marité Arleth, Lise Nanoscale Adv Chemistry Supported lipid bilayers (SLBs) are commonly used as model systems mimicking biological membranes. Recently, we reported a new method to produce SLBs with incorporated membrane proteins, which is based on the application of peptide discs [Luchini et al., Analytical Chemistry, 2020, 92, 1081–1088]. Peptide discs are small discoidal particles composed of a lipid core and an outer belt of self-assembled 18A peptides. SLBs including membrane proteins can be formed by depositing the peptide discs on a solid support and subsequently removing the peptide by buffer rinsing. Here, we introduce a new variant of the 18A peptide, named dark peptide (d18A). d18A exhibits UV absorption at 214 nm, whereas the absorption at 280 nm is negligible. This improves sample preparation as it enables a direct quantification of the membrane protein concentration in the peptide discs by measuring UV absorption at 280 nm. We describe the application of the peptide discs prepared with d18A (dark peptide discs) to produce SLBs with a membrane protein, synaptobrevin 2 (VAMP2). The collected data showed the successful formation of SLBs with high surface coverage and incorporation of VAMP2 in a single orientation with the extramembrane domain exposed towards the bulk solvent. Compared to 18A, we found that d18A was more efficiently removed from the SLB. Our data confirmed the structural organisation of VAMP2 as including both α-helical and β-sheet secondary structure. We further verified the orientation of VAMP2 in the SLBs by characterising the binding of VAMP2 with α-synuclein. These results point at the produced SLBs as relevant membrane models for biophysical studies as well as nanostructured biomaterials. RSC 2022-09-12 /pmc/articles/PMC9595196/ /pubmed/36341300 http://dx.doi.org/10.1039/d2na00384h Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Luchini, Alessandra
Tidemand, Frederik Grønbæk
Johansen, Nicolai Tidemand
Sebastiani, Federica
Corucci, Giacomo
Fragneto, Giovanna
Cárdenas, Marité
Arleth, Lise
Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2)
title Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2)
title_full Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2)
title_fullStr Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2)
title_full_unstemmed Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2)
title_short Dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (VAMP2)
title_sort dark peptide discs for the investigation of membrane proteins in supported lipid bilayers: the case of synaptobrevin 2 (vamp2)
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9595196/
https://www.ncbi.nlm.nih.gov/pubmed/36341300
http://dx.doi.org/10.1039/d2na00384h
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