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Title: Engineering phosphatidylinositol-4,5-bisphosphate model membranes enriched in endocytic cargo: a neutron reflectometry, AFM and QCM-D structural study
Author: Pereira, Daniel
Santamaria, Andreas
Pawar, Nisha
Carrascosa-Tejedor, Javier
Sardo, Mariana
Mafra, Luís
Guzmán, Eduardo
Owen, David J.
Zaccai, Nathan R.
Maestro, Armando
Marín-Montesinos, Ildefonso
Keywords: Neutron reflectometry
Quartz crystal microbalance with dissipation monitoring
Membrane biomimetics
Issue Date: Jul-2023
Publisher: Elsevier
Abstract: The combination of in vitro models of biological membranes based on solid-supported lipid bilayers (SLBs) and of surface sensitive techniques, such as neutron reflectometry (NR), atomic force microscopy (AFM) and quartz crystal microbalance with dissipation monitoring (QCM-D), is well suited to provide quantitative information about molecular level interactions and lipid spatial distributions. In this work, cellular plasma membranes have been mimicked by designing complex SLB, containing phosphatidylinositol 4,5-bisphosphate (PtdIns4,5P2) lipids as well as incorporating synthetic lipo-peptides that simulate the cytoplasmic tails of transmembrane proteins. The QCM-D results revealed that the adsorption and fusion kinetics of PtdIns4,5P2 are highly dependent of Mg2+. Additionally, it was shown that increasing concentrations of PtdIns4,5P2 leads to the formation of SLBs with higher homogeneity. The presence of PtdIns4,5P2 clusters was visualized by AFM. NR provided important insights about the structural organization of the various components within the SLB, highlighting that the leaflet symmetry of these SLBs is broken by the presence of CD4-derived cargo peptides. Finally, we foresee our study to be a starting point for more sophisticated in vitro models of biological membranes with the incorporation of inositol phospholipids and synthetic endocytic motifs.
Peer review: yes
DOI: 10.1016/j.colsurfb.2023.113341
ISSN: 0927-7765
Appears in Collections:CICECO - Artigos
DQ - Artigos

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