| 1. |
- Berts, Ida, 1984-, et al.
(author)
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Controlling adsorption of albumin with hyaluronan on silica surfaces and sulfonated latex particles
- 2017
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In: Journal of Colloid and Interface Science. - : Elsevier BV. - 0021-9797 .- 1095-7103. ; 504, s. 315-324
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Journal article (peer-reviewed)abstract
- Polysaccharides are known to modify binding of proteins at interfaces and this paper describes studies of these interactions and how they are modified by pH. Specifically, the adsorption of human serum albumin on to polystyrene latex and to silica is described, focusing on how this is affected by hyaluronan. Experiments were designed to test how such binding might be modified under relevant physiological conditions. Changes in adsorption of albumin alone and the co-adsorption of albumin and hyaluronan are driven by electrostatic interactions. Multilayer binding is found to be regulated by the pH of the solution and the molecular mass and concentration of hyaluronan. Highest adsorption was observed at pH below 4.8 and for low molecular mass hyaluronan (<= 150 kDa) at concentrations above 2 mg ml(-1). On silica with grafted hyaluronan, albumin absorption is reversed by changes in solvent pH due to their strong electrostatic attraction. Albumin physisorbed on silica surfaces is also rinsed away with dilute hyaluronan solution at pH 4.8. The results demonstrate that the protein adsorption can be controlled both by changes of pH and by interaction with other biological macromolecules.
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| 2. |
- de Ghellinck, Alexis, et al.
(author)
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Lipid polyunsaturation determines the extent of membrane structural changes induced by Amphotericin B in Pichia pastoris yeast
- 2015
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In: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 0005-2736. ; 1848:10, s. 2317-2325
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Journal article (peer-reviewed)abstract
- The activity of the potent but highly toxic antifungal drug Amphotericin B (AmB), used intravenously to treat systemic fungal and parasitic infections, is widely accepted to result from its specific interaction with the fungal sterol ergosterol. While the effect of sterols on AmB activity has been intensely investigated, the role of membrane phospholipid composition has largely been ignored, and structural studies of native membranes have been hampered by their complex and disordered nature. We show for the first time that the structure of fungal membranes derived from Pichia pastoris yeast depends on the degree of lipid polyunsaturation, which has an impact on the structural consequences of AmB activity. AmB inserts in yeast membranes even in the absence of ergosterol, and forms an extra-membraneous layer whose thickness is resolved to be 4-5 nm. In ergosterol-containing membranes, AmB insertion is accompanied by ergosterol extraction into this layer. The AmB-sponge mediated depletion of ergosterol from P. pastoris membranes gives rise to a significant membrane thinning effect that depends on the degree of lipid polyunsaturation. The resulting hydrophobic mismatch is likely to interfere with a much broader range of membrane protein functions than those directly involving ergosterol, and suggests that polyunsaturated lipids could boost the efficiency of AmB. Furthermore, a low degree of lipid polyunsaturation leads to least AmB insertion and may protect host cells against the toxic effects of AmB. These results provide a new framework based on lipid composition and membrane structure through which we can understand its antifungal action and develop better treatments. (C) 2015 Elsevier B.V. All rights reserved.
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| 3. |
- Fragneto, Giovanna, et al.
(author)
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Neutrons and model membranes : Moving towards complexity
- 2018
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In: Current Opinion in Colloid and Interface Science. - : Elsevier BV. - 1359-0294. ; 38, s. 108-121
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Research review (peer-reviewed)abstract
- Cells, the basic units of living organisms, are well delineated and separated from the external environment by membranes. Capable of both enclosing the cellular constituents and allowing exchanges with the outside world, these membranes are only a few nanometers thick. All the membranes in a human body cover an area of a few hectares, but account for only a small part of our mass. To study the dynamics and function of these amazing objects, physicists first seek to understand their structure. This involves experiments on model systems, simpler and better controlled than real membranes, and can profit from a probe that is able to access different scales of size and time: thermal neutrons. Since the pioneering work in the seventies on cell membrane structure by neutron scattering, developments driven by constantly improving neutron instrumentation, coupled with development of measurement and analysis methods, have involved both the optimization of samples towards more biologically relevant model systems and include the use of more complex lipid mixtures up to natural extracts. This review does not have the ambition to address the large number of contributions from all the groups working in this area in research laboratories and neutron facilities. It gives an update on some studies in the field carried out mainly by the authors and collaborators.
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| 4. |
- Luchini, Alessandra, et al.
(author)
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Neutron Reflectometry reveals the interaction between functionalized SPIONs and the surface of lipid bilayers
- 2017
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In: Colloids and Surfaces B. - : ELSEVIER SCIENCE BV. - 0927-7765 .- 1873-4367. ; 151, s. 76-87
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Journal article (peer-reviewed)abstract
- The safe application of nanotechnology devices in biomedicine requires fundamental understanding on how they interact with and affect the different components of biological systems. In this respect, the cellular membrane, the cell envelope, certainly represents an important target or barrier for nanosystems. Here we report on the interaction between functionalized SuperParamagnetic Iron Oxide Nanoparticles (SPIONs), promising contrast agents for Magnetic Resonance Imaging (MRI), and lipid bilayers that mimic the plasma membrane. Neutron Reflectometry, supported by Quartz Crystal Microbalance with Dissipation monitoring (QCM-D) experiments, was used to characterize this interaction by varying both SPION coating and lipid bilayer composition. In particular, the interaction of two different SPIONs, functionalized with a cationic surfactant and a zwitterionic phospholipid, and lipid bilayers, containing different amount of cholesterol, were compared. The obtained results were further validated by Dynamic Light Scattering (DLS) measurements and Cryogenic Transmission Electron Microscopy (Cryo-TEM) images. None of the investigated functionalized SPIONs were found to disrupt the lipid membrane. However, in all case we observed the attachment of the functionalized SPIONs onto the surface of the bilayers, which was affected by the bilayer rigidity, i.e. the cholesterol concentration.
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| 5. |
- Luchini, Alessandra, et al.
(author)
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The impact of deuteration on natural and synthetic lipids : A neutron diffraction study
- 2018
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In: Colloids and Surfaces B: Biointerfaces. - : Elsevier BV. - 0927-7765. ; 168, s. 126-133
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Journal article (peer-reviewed)abstract
- The structural investigation of cellular membranes requires access to model systems where the molecular complexity is representative of the cellular environment and that allow for the exploitation of structural techniques. Neutron scattering, and in particular neutron diffraction can provide unique and detailed information on the structure of lipid membranes. However, deuterated samples are desirable to fully exploit this powerful method. Recently, the extraction of lipids from microorganisms grown in deuterated media was demonstrated to be both an attracting route to obtain complex lipid mixtures resembling the composition of natural membranes, and to producing deuterated molecules in a very convenient way. A full characterization of these deuterated extracts is hence pivotal for their use in building up model membrane systems. Here we report the structural characterization of lipid extracts obtained from Pichia pastoris by means of neutron diffraction measurements. In particular, we compare the structure of membranes extracted from yeast cells grown in a standard culture medium and in a corresponding deuterated culture medium. The results show that the different molecular composition of the deuterated and protiated lipid extracts induce different structural organization of the lipid membranes. In addition, we compare these membranes composed of extracted yeast lipids with stacked bilayers prepared from synthetic lipid mixtures.
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| 6. |
- Montis, Costanza, et al.
(author)
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Nucleolipid bilayers: A quartz crystal microbalance and neutron reflectometry study.
- 2016
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In: Colloids and Surfaces B: Biointerfaces. - : Elsevier BV. - 1873-4367 .- 0927-7765. ; 137:Online 22 July 2015, s. 203-213
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Journal article (peer-reviewed)abstract
- POP-Ade (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidyladenosine) is a biocompatible anionic nucleolipid with the DNA nucleoside, Adenosine, in the polar headgroup. We have studied the affinity of nucleic acids of different contour length, composition and structure toward supported lipid bilayers (SLB) composed of POP-Ade mixed with the zwitterionic phospholipid POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) using quartz crystal microbalance with dissipation monitoring (QCM-D) and neutron reflectometry (NR). In order to highlight the specificity of the nucleic acid interaction, the results were compared with data obtained for SLB containing the anionic phospholipid POPG (1-palmitoyl-2-oleoyl-sn-phosphatidyl-glycerol) replacing POP-Ade. Our results demonstrate that the presence of a nucleobase headgroup provides the bilayers with the ability to bind single stranded nucleic acids in a selective fashion, according to a Watson-Crick pattern. In addition the interaction with double stranded nucleic acids was strengthened. Overall, these findings represent fundamental information for the design of biocompatible DNA vectors with DNA-RNA-based amphiphiles.
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