Design and use of model membranes to study biomolecular interactions using complementary surface-sensitive techniques

Clifton, Luke A.; Campbell, Richard A.; Sebastiani, Federica; Campos‐Terán, José; Gonzalez-Martinez, Juan Francisco; Björklund, Sebastian; Sotres, Javier; Cárdenas, Marité

doi:10.1016/j.cis.2020.102118

Cited by 78 publications

(85 citation statements)

References 207 publications

(269 reference statements)

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“…NR also allows to simultaneously resolve potential lipid removal as well as peptide insertion into partly deuterated supported lipid bilayers (SLBs). [31][32][33][34][35][36][37][38] In an earlier work, we showed that NR results can be directly compared to results from detailed modelling of small angle X-ray scattering (SAXS) data on monomeric peptide lipid bilayer using SLBs or unilamellar vesicles respectively. 31 For supramolecular nanobers in particular, NR has an advantage over bulk methods since it lacks 3D orientation averaging and enables precise structural determination of complex MDP-membrane structures.…”

Section: Introductionmentioning

confidence: 99%

Lipid membrane interactions of self-assembling antimicrobial nanofibers: effect of PEGylation

et al. 2020

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Section: Introductionmentioning

confidence: 99%

Lipid membrane interactions of self-assembling antimicrobial nanofibers: effect of PEGylation

et al. 2020

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“…In this study, we firstly employ quantitative single particle microscopy [ 26 , 27 ], and arrays of surface tethered liposomes as model cell membranes [ 28 , 29 ] to observe directly several successive docking events of SL assemblies on individual nanoscale liposomes (see Figure 1 ) and provide a quantitative understanding of how membrane charge and acidic pH may regulate these interactions. Utilizing a biophysical system such as liposomes enables a deconvolution of regulatory effects (i.e., pH and surface charge) as well as the observation of several successive events that remain masked in contemporary measurements and thus provide insights into the underlying mechanism of interactions.…”

Section: Introductionmentioning

confidence: 99%

Direct Observation of Sophorolipid Micelle Docking in Model Membranes and Cells by Single Particle Studies Reveals Optimal Fusion Conditions

2020

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Sophorolipids (SLs) are naturally produced glycolipids that acts as drug delivery for a spectrum of biomedical applications, including as an antibacterial antifungal and anticancer agent, where they induce apoptosis selectively in cancerous cells. Despite their utility, the mechanisms underlying their membrane interactions, and consequently cell entry, remains unknown. Here, we combined a single liposome assay to observe directly and quantify the kinetics of interaction of SL micelles with model membrane systems, and single particle studies on live cells to record their interaction with cell membranes and their cytotoxicity. Our single particle readouts revealed several repetitive docking events on individual liposomes and quantified how pH and membrane charges, which are known to vary in cancer cells, affect the docking of SL micelles on model membranes. Docking of sophorolipids micelles was found to be optimal at pH 6.5 and for membranes with -5% negatively charge lipids. Single particle studies on mammalian cells reveled a two-fold increased interaction on Hela cells as compared to HEK-293 cells. This is in line with our cell viability readouts recording an approximate two-fold increased cytotoxicity by SLs interactions for Hela cells as compared to HEK-293 cells. The combined in vitro and cell assays thus support the increased cytotoxicity of SLs on cancer cells to originate from optimal charge and pH interactions between membranes and SL assemblies. We anticipate studies combining quantitative single particle studies on model membranes and live cell may reveal hitherto unknown molecular insights on the interactions of sophorolipid and additional nanocarriers mechanism.

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“…The presence of membrane domains enriched in cholesterol and sphingolipids was also reproduced in biomimetic lipid membranes and resulted in having a strong impact on the interaction with the Aβ peptide [101]. In this context, the characterization by X-ray diffraction of supported lipid membranes made of dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphoserine (DMPS), and cholesterol (at 30 mol%) demonstrated the effect of the membrane lipid composition in modulating the interactions with Aβ(1-42) and Aβ (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35) fragments [102]. Additional recent studies on Aβ-membrane interaction used vesicles and supported lipid bilayers with a complex lipid composition, including raft lipids such as cholesterol and sphingomyelin (SM), but also polyunsaturated fatty acids (PUFA) (better known as omega-3 lipids), and showed the central role of omega-3 lipids in favoring a deeper internalization of the peptide among the lipid acyl chains and, consequently, hindering its pathogenic self-aggregation [103,104].…”

Section: Protein-lipid Interactionsmentioning

confidence: 92%

“…Indeed, scattering techniques, including neutrons, X-ray, and light as probes, spectroscopy techniques, e.g., fluorescence spectroscopy, NMR, electron paramagnetic resonance (EPR) spectroscopy, and calorimetry are typically used for the investigation of lipid membranes in solution [27][28][29]. On the other hand, surface sensitive techniques such as neutron and X-ray reflectometry and diffraction, Langmuir isotherms, attenuated total reflectance-FTIR, and microscopy techniques, e.g., electron microscopy, fluorescence microscopy, atomic force microscopy, and Brewster angle microscopy, are suitable for the characterization of lipid membranes on surfaces [29,30]. In this section, we discuss the main characteristics of the two types of biomimetic lipid membranes together with a brief description of their respective sample preparation protocols.…”

Section: Design Of Biomimetic Lipid Membranesmentioning

confidence: 99%

Mimicking the Mammalian Plasma Membrane: An Overview of Lipid Membrane Models for Biophysical Studies

Luchini

Vitiello

2020

Biomimetics

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Cell membranes are very complex biological systems including a large variety of lipids and proteins. Therefore, they are difficult to extract and directly investigate with biophysical methods. For many decades, the characterization of simpler biomimetic lipid membranes, which contain only a few lipid species, provided important physico-chemical information on the most abundant lipid species in cell membranes. These studies described physical and chemical properties that are most likely similar to those of real cell membranes. Indeed, biomimetic lipid membranes can be easily prepared in the lab and are compatible with multiple biophysical techniques. Lipid phase transitions, the bilayer structure, the impact of cholesterol on the structure and dynamics of lipid bilayers, and the selective recognition of target lipids by proteins, peptides, and drugs are all examples of the detailed information about cell membranes obtained by the investigation of biomimetic lipid membranes. This review focuses specifically on the advances that were achieved during the last decade in the field of biomimetic lipid membranes mimicking the mammalian plasma membrane. In particular, we provide a description of the most common types of lipid membrane models used for biophysical characterization, i.e., lipid membranes in solution and on surfaces, as well as recent examples of their applications for the investigation of protein-lipid and drug-lipid interactions. Altogether, promising directions for future developments of biomimetic lipid membranes are the further implementation of natural lipid mixtures for the development of more biologically relevant lipid membranes, as well as the development of sample preparation protocols that enable the incorporation of membrane proteins in the biomimetic lipid membranes.

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Design and use of model membranes to study biomolecular interactions using complementary surface-sensitive techniques

Cited by 78 publications

References 207 publications

Lipid membrane interactions of self-assembling antimicrobial nanofibers: effect of PEGylation

Lipid membrane interactions of self-assembling antimicrobial nanofibers: effect of PEGylation

Direct Observation of Sophorolipid Micelle Docking in Model Membranes and Cells by Single Particle Studies Reveals Optimal Fusion Conditions

Mimicking the Mammalian Plasma Membrane: An Overview of Lipid Membrane Models for Biophysical Studies

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