The complexity of the biological membranes restricts their direct investigation at the nanoscale. Lipid bilayer membranes have been developed as a model of biological membranes in order to allow the interaction and insertion of peptides and membrane proteins in a functional manner. Promising models have been developed in the past two decades and tethered bilayer design traduces constant improvement of membrane models. The formation of protein free solid tethered membranes can be achieved by direct vesicle fusion, Langmuir-Blodgett, Langmuir-Schaffer transfers, self assembly of various building blocks such as thiol on gold, silane on quartz, grafting of polymers, as well as ligand receptor recognition. In this review, the current state of different tethered bilayer membrane will be described. We will focus on critical analysis of the main advantages/drawbacks of each kind of model construction and their ability to allow protein incorporation in non-denaturing conditions. Some of the current drawbacks encountered in these biomimetic models can be overcome using an innovative tethered bilayer design based on a reliable and fast formation method. The successful protein incorporation of the Adenylate Cyclase produced by Bordetella pertussis and the voltage dependent anion channel (VDAC) was demonstrated on this model.
Over the past two decades, numerous types of nanoparticles (NPs) have been developed for medical applications; however only a few nanomedicines are actually available on the market. One reason is the lack of understanding and data concerning the NP fate and their behavior upon contact with biological media and cell membranes. Biomimetic membrane models are interesting tools to approach and understand NPs-cell membrane interactions. The use of these models permits one to control physical and chemical parameters and to rapidly compare membrane types and the influence of different media conditions. The interactions between NPs and cell membranes can be qualified and quantified using analytical and modeling methods. In this review, the major studies concerning NPs-cell membrane models and associated methods are described. The advantages and drawbacks for each method are compared for the different models. The key mechanisms of interactions between NPs and cell membranes are revealed using cell membrane models and are interrogated in comparison with the NP behavior in cellulo or in vivo. Investigating the interactions between NPs and cell membrane models is now proposed as an intermediate step between physicochemical characterization of NPs and biological assays.
Background:The translocation of the Bordetella pertussis CyaA toxin across membrane is still poorly understood. Results: A membrane-active peptide isolated from the CyaA toxin is characterized by biophysical approaches.
Conclusion:The ␣-helical peptide is inserted in plane and induces membrane permeabilization. Significance: The membrane-destabilizing activity of this peptide may assist the initial steps of the CyaA translocation process.
In this work, two different types of supported biomimetic membranes were designed to study the membrane binding properties of two different proteins that both interact with cellular membranes in a calcium-dependent manner. The first one, neurocalcin, is a member of a subfamilly of EF-hand calcium-binding proteins that exhibit a calcium-myristoyl switch. The second protein is a bacterial toxin, the adenylate cyclase produced by Bordetella pertussis, the causative agent of whooping cough. The biomimetic membranes constructed in this study were either hybrid bilayer membranes or polymer-tethered membranes. Hemimembrane formation was obtained in two steps: a monolayer of 1-octadecanethiol or octadecyltrichlorosilane was self-assembled on top of the gold or glass surface, respectively, and then the egg-phosphatidyl choline (PC) vesicle fused on the hydrophobic alkyl layer. Polymer-tethered membranes on solid support were obtained using N-hydroxysuccinimide (NHS)-terminated-poly(ethyleneglycol) (PEG)-phospholipids as anchoring molecules. Egg-PC/1,2-distearoyl-sn-glycero-3-phospho-ethanolamine-poly(ethyleneglycol)-N-hydroxy-succinimide (DSPE-PEG-NHS) mixture liposomes were injected on the top of an amine grafted surface (cysteamine-coated gold or silanized glass); vesicles were linked to the surface and disrupted, leading to the formation of a bilayer. The biomimetic membrane constructions were followed by surface plasmon spectroscopy, while membrane fluidity and continuity were observed by fluorescence microscopy. Protein/membrane binding properties were determined by resonance surface plasmon measurements. The tethered bilayer, designed here, is very versatile as it can be adapted easily to different types of support. The results demonstrate the potentialities of such polymer-tethered artificial membranes for the study of proteins that insert into biological membranes such as toxins and/or integral membrane proteins.
Magnetic mesoporous silica nanoparticles (M-MSNs) represent promising targeting tools for theranostics. Engineering the interaction of nanoparticles (NPs) with biological systems requires an understanding of protein corona formation around the nanoparticles as this drives the biological fate of nanocarriers. We investigated the behavior of proteins in contact with M-MSNs by high-throughput comparative proteomics, using human and bovine sera as biological fluids, in order to assess the adsorption dynamics of proteins in these media. Using system biology tools, and especially protein-protein interaction databases, we demonstrated how the protein network builds up within the corona over the course of the experiment. Based on these results, we introduce and discuss the role of the "corona interactome" as an important factor influencing protein corona evolution. The concept of the "corona interactome" is an original methodology which could be generalized to all NP candidates. Based on this, pre-coating nanocarriers with specific proteins presenting minimal interactions with opsonins might provide them with properties such as stealth.
Significance
Many bacterial toxins can cross biological membranes to reach the cytosol of mammalian cells, although how they pass through a lipid bilayer remains largely unknown.
Bordetella pertussis
adenylate cyclase (CyaA) toxin delivers its catalytic domain directly across the cell membrane. To characterize this unique translocation process, we designed an in vitro assay based on a tethered lipid bilayer assembled over a biosensor surface derivatized with calmodulin, a natural activator of the toxin. CyaA activation by calmodulin provided a highly sensitive readout for toxin translocation across the bilayer. CyaA translocation was calcium- and membrane potential-dependent but independent of any additional eukaryotic protein. This biomimetic membrane will permit in vitro studies of protein translocation in precisely defined conditions.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.