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See detailValorisation d’un co-produit de la production du bio-éthanol comme substrat de fermentation pour la production de nouvelles formes de biosurfactants plus actives
Lins, Laurence ULiege; Deleu, Magali ULiege; Jacques, Philippe ULiege

Conference (2021, June)

Les surfactants d'origine biologique suscitent un intérêt croissant dans différents domaines applicatifs tels que la détergence, la cosmétique, le traitement des sols pollués ou encore la dispersion des ... [more ▼]

Les surfactants d'origine biologique suscitent un intérêt croissant dans différents domaines applicatifs tels que la détergence, la cosmétique, le traitement des sols pollués ou encore la dispersion des marées noires en raison de leurs propriétés uniques telles que leur spécificité, leur biodégradabilité et leur toxicité potentiellement plus faible. Pour ces applications, l’obstacle majeur reste le coût de production. Dans le cadre du projet européen BestBioSurf, 7 laboratoires et 3 entreprises ont mis ensemble leurs compétences pour développer de nouveaux biosurfactants aux propriétés de détergence accrue ainsi qu’un procédé de production permettant d’en réduire le prix de revient. En se basant sur les propriétés de biosurfactants d’origine microbienne, des outils informatiques de prédiction des relations structure-fonction ont été utilisés pour prédire la(les) structure(s) du biosurfactant la(les) plus optimale(s) en termes de propriétés physico-chimiques. Les souches productrices des biosurfactants les plus efficaces ont été cultivées sur un co-produit de la production du bio-éthanol à partir de céréales. Ce co-produit généré lors de la distillation répond à de nombreux critères essentiels pour être utilisé à cette fin : la présence de tous les substrats nécessaires à la croissance des micro-organismes concernés et à la production des biosurfactants, l’absence d’inhibiteurs, un tonnage important et un coût extrêmement faible. [less ▲]

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See detailTruncated YY1 interacts with BASP1 through a 339KLK341 motif in YY1 and suppresses vascular smooth muscle cell growth and intimal hyperplasia after vascular injury.
Santiago, Fernando S.; Li, Yue; Zhong, Ling et al

in Cardiovascular Research (2021)

AIMS: In-stent restenosis and late stent thrombosis are complications associated with the use of metallic and drug-coated stents. Strategies that inhibit vascular smooth muscle cell (SMC) proliferation ... [more ▼]

AIMS: In-stent restenosis and late stent thrombosis are complications associated with the use of metallic and drug-coated stents. Strategies that inhibit vascular smooth muscle cell (SMC) proliferation without affecting endothelial cell (EC) growth would be helpful in reducing complications arising from percutaneous interventions. Our group previously showed that the forced expression of the injury-inducible zinc finger (ZNF) transcription factor, yin yang-1 (YY1) comprising 414 residues inhibits neointima formation in carotid arteries of rabbits and rats. YY1 inhibits SMC proliferation without affecting EC growth. Identifying a shorter version of YY1 retaining cell-selective inhibition would make it more amenable for potential use as a gene therapeutic agent. METHODS AND RESULTS: We dissected YY1 into a range of shorter fragments (YY1A-D, YY1Δ) and found that the first two ZNFs in YY1 (construct YY1B, spanning 52 residues) repressed SMC proliferation. Receptor Binding Domain analysis predicts a three residue (339KLK341) interaction domain. Mutation of 339KLK341 to 339AAA341 in YY1B (called YY1Bm) abrogated YY1B's ability to inhibit SMC but not EC proliferation and migration. Incubation of recombinant GST-YY1B and GST-YY1Bm with SMC lysates followed by precipitation with glutathione-agarose beads and mass spectrometric analysis identified a novel interaction between YY1B and BASP1. Overexpression of BASP1, like YY1, inhibited SMC but not EC proliferation and migration. BASP1 siRNA partially rescued SMC from growth inhibition by YY1B. In the rat carotid balloon injury model, adenoviral overexpression of YY1B, like full-length YY1, reduced neointima formation, whereas YY1Bm had no such effect. CD31 immunostaining suggested YY1B could increase re-endothelialization in a 339KLK341-dependent manner. CONCLUSIONS: These studies identify a truncated form of YY1 (YY1B) that can interact with BASP1 and inhibits SMC proliferation, migration and intimal hyperplasia after balloon injury of rat carotid arteries as effectively as full length YY1. We demonstrate the therapeutic potential of YY1B in vascular proliferative disease. [less ▲]

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See detailBiophysical analysis of the plant-specific GIPC sphingolipids reveals multiple modes of membrane regulation.
Mamode Cassim, Adiilah; Navon, Yotam; Gao, Yuan ULiege et al

in The Journal of biological chemistry (2021), 296

The plant plasma membrane (PM) is an essential barrier between the cell and the external environment, controlling signal perception and transmission. It consists of an asymmetrical lipid bilayer made up ... [more ▼]

The plant plasma membrane (PM) is an essential barrier between the cell and the external environment, controlling signal perception and transmission. It consists of an asymmetrical lipid bilayer made up of three different lipid classes: sphingolipids, sterols and phospholipids. The Glycosyl Inositol Phosphoryl Ceramides (GIPCs), representing up to 40% of total sphingolipids, are assumed to be almost exclusively in the outer leaflet of the PM. However, their biological role and properties are poorly defined. In this study, we investigated the role of GIPCs in membrane organization. Since GIPCs are not commercially available, we developed a protocol to extract and isolate GIPC-enriched fractions from eudicots (cauliflower and tobacco) and monocots (leek and rice). Lipidomic analysis confirmed the presence of trihydroxylated long chain bases and 2-hydroxylated very long chain fatty acids up to 26 carbon atoms. The glycan head groups of the GIPCs from monocots and dicots were analyzed by GC-MS, revealing different sugar moieties. Multiple biophysics tools, namely Langmuir monolayer, ζ-Potential, light scattering, neutron reflectivity, solid state 2H-NMR and molecular modelling, were used to investigate the physical properties of the GIPCs, as well as their interaction with free and conjugated phytosterols. We showed that GIPCs increase the thickness and electronegativity of model membranes, interact differentially with the different phytosterols species and regulate the gel-to-fluid phase transition during temperature variations. These results unveil the multiple roles played by GIPCs in the plant plasma membrane. [less ▲]

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See detailLIMONADA: A database dedicated to the simulation of biological membranes.
Crowet, Jean-Marc; Buchoux, Sébastien; Belloy, Nicolas et al

in Journal of computational chemistry (2021), 42(14), 1028-1033

Cellular membranes are composed of a wide diversity of lipid species in varying proportions and these compositions are representative of the organism, cellular type and organelle to which they belong ... [more ▼]

Cellular membranes are composed of a wide diversity of lipid species in varying proportions and these compositions are representative of the organism, cellular type and organelle to which they belong. Because models of these molecular systems simulated by MD steadily gain in size and complexity, they are increasingly representative of specific compositions and behaviors of biological membranes. Due to the number of lipid species involved, of force fields and topologies and because of the complexity of membrane objects that have been simulated, LIMONADA has been developed as an open database allowing to handle the various aspects of lipid membrane simulation. LIMONADA presents published membrane patches with their simulation files and the cellular membrane it models. Their compositions are then detailed based on the lipid identification from LIPID MAPS database plus the lipid topologies and the force field used. LIMONADA is freely accessible on the web at https://limonada.univ-reims.fr/. [less ▲]

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See detailA compartmentalized microsystem helps understanding the uptake of benzo[a]pyrene by fungi during soil bioremediation processes.
Baranger, Claire; Pezron, Isabelle; Lins, Laurence ULiege et al

in The Science of the total environment (2021), 784

Hydrophobic organic soil contaminants such as polycyclic aromatic hydrocarbons (PAH) are poorly mobile in the aqueous phase and tend to sorb to the soil matrix, resulting in low bioavailability. Some ... [more ▼]

Hydrophobic organic soil contaminants such as polycyclic aromatic hydrocarbons (PAH) are poorly mobile in the aqueous phase and tend to sorb to the soil matrix, resulting in low bioavailability. Some filamentous fungi are efficient in degrading this kind of pollutants. However, the mechanism of mobilization of hydrophobic compounds by non-motile microorganisms such as filamentous fungi needs investigations to improve pollutant bioavailability and bioremediation efficiency. Usual homogeneous media for microbial growth in the lab are poorly suited to model the soil, which is a compartmentalized and heterogeneous habitat. A microfluidic device was designed to implement a compartmentalization of the fungal inoculum and the source of the pollutant benzo[a]pyrene (BaP) as a deposit of solid crystals in order to gain a further insight into the mechanisms involved in the access to the contaminant and its uptake in soils. Thus in this device, two chambers are connected by an array of parallel microchannels that are wide enough to allow individual hyphae to grow through them. Macro-cultures of Talaromyces helicus in direct contact with BaP have shown its uptake and intracellular storage in lipid bodies despite the low propensity of BaP to cross aqueous phases as shown by simulation. Observations of T. helicus in the microfluidic device through laser scanning confocal microscopy indicate preferential uptake of BaP at a close range and through contact with the cell wall. However faint staining of some hyphae before contact with the deposit also suggests an extracellular transport phenomenon. Macro-culture filtrates analyses have shown that T. helicus releases extracellular non-lipidic surface-active compounds able to lower the surface tension of culture filtrates to 49.4 mN/m. Thus, these results highlight the significance of active mechanisms to reach hydrophobic contaminants before their uptake by filamentous fungi in compartmentalized micro-environments and the potential to improve them through biostimulation approaches for soil mycoremediation. [less ▲]

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See detailThe Surfactin-Like Lipopeptides From Bacillus spp.: Natural Biodiversity and Synthetic Biology for a Broader Application Range
Théatre, Ariane ULiege; Cano-Prieto, Carolina; Bartolini, Marco et al

in Frontiers in Bioengineering and Biotechnology (2021), 9

Surfactin is a lipoheptapeptide produced by several Bacillus species and identified for the first time in 1969. At first, the biosynthesis of this remarkable biosurfactant was described in this review ... [more ▼]

Surfactin is a lipoheptapeptide produced by several Bacillus species and identified for the first time in 1969. At first, the biosynthesis of this remarkable biosurfactant was described in this review. The peptide moiety of the surfactin is synthesized using huge multienzymatic proteins called NonRibosomal Peptide Synthetases. This mechanism is responsible for the peptide biodiversity of the members of the surfactin family. In addition, on the fatty acid side, fifteen different isoforms (from C12 to C17) can be incorporated so increasing the number of the surfactin-like biomolecules. The review also highlights the last development in metabolic modeling and engineering and in synthetic biology to direct surfactin biosynthesis but also to generate novel derivatives. This large set of different biomolecules leads to a broad spectrum of physico-chemical properties and biological activities. The last parts of the review summarized the numerous studies related to the production processes optimization as well as the approaches developed to increase the surfactin productivity of Bacillus cells taking into account the different steps of its biosynthesis from gene transcription to surfactin degradation in the culture medium. [less ▲]

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See detailLinolenic fatty acid hydroperoxide acts as biocide on plant pathogenic bacteria: biophysical investigation of the mode of action
Deboever, Estelle ULiege; Lins, Laurence ULiege; Ongena, Marc ULiege et al

in Bioorganic Chemistry (2020), 100

Fatty acid hydroperoxides (HPO) are free phyto-oxylipins known for their crucial role as signalling molecules during plant defense mechanisms. They were also demonstrated to have direct biocidal ... [more ▼]

Fatty acid hydroperoxides (HPO) are free phyto-oxylipins known for their crucial role as signalling molecules during plant defense mechanisms. They were also demonstrated to have direct biocidal activities against plant pathogens including gram negative bacteria. In the present work, the biocidal effect of one linolenic fatty acid hydroperoxide, namely 13-HPOT has been investigated on three plant pathogen gram negative bacteria: Pectobacterium carotovorum, Pseudomonas syringae and Xanthomonas translucens. We showed that 13-HPOT has a strong dose response effect on those phytopathogens. In a second part, the molecular mechanism behind the antibacterial effect of 13-HPOT was investigated at a molecular level using an integrative biophysical approach combining in vitro and in silico methods. Since other antimicrobial amphiphilic molecules have been shown to target the lipid membrane of the organisms they act on, we focused our study on the interaction of 13-HPOT with biomimetic membranes. In a first step, we hypothesized that the inner membrane of the bacteria was the main site of action of 13-HPOT and hence we used lipids representative of this membrane to form our models. Our results indicated that 13-HPOT can interact with the lipid representative of the inner bacterial plasma membrane. A strong membrane insertion is suggested but no major permeabilization of the membrane is observed. Phosphatidylethanolamine (PE) and cardiolipin (CL), present in the bacterial plasma membrane, appear to play important roles in this interaction. We suggest that the mode of action of 13-HPOT should involve either subtle changes in membrane properties, such as its lateral organization and distribution, and/or interactions with membrane proteins. [less ▲]

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See detailMolecular understanding of the interactions between modified surfactins and bacterial membranes
Lins, Laurence ULiege; Deleu, Magali ULiege; Phalip, Vincent

Scientific conference (2020, May)

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See detailMolecular biophysics as a tool to investigate the bioherbicide effect of essential oils related to their interaction with plant plasma membrane
Lins, Laurence ULiege; De Clerck, Caroline ULiege; Fauconnier, Marie-Laure ULiege et al

Conference (2020, February)

Essential oils (EOs) are used in an increasingly number of sectors like medicine, cosmetics, food industry and more recently in agronomy. In agronomy, EOs are used as bio-pesticides for their insecticidal ... [more ▼]

Essential oils (EOs) are used in an increasingly number of sectors like medicine, cosmetics, food industry and more recently in agronomy. In agronomy, EOs are used as bio-pesticides for their insecticidal, antifungal or bactericidal effects but also as bio-herbicides. Owing to the current attraction for natural products, a better understanding of their mode of biological action for new and optimal applications is of importance. It has been shown that EOs antimicrobial activity, quite well described in the literature, is at least partly due to their interaction with the plasma membrane. They notably change the lipid composition, altering fluidity, leading to various effects which can induce cell lysis, apoptosis or necrosis. Citronellol, a major compound of lemongrass EO was notably shown to have antifungal activities by changing the membrane composition and inhibiting cell growth. Cinnamaldehyde (cinnamon EO) has been reported to have a broad spectrum of antibacterial activity, notably by affecting cell morphology, membrane integrity, permeability and composition. We are currently working on the development of a bioherbicide made from Cinnamomum zeylanicum Blume (cinnamon) and Cymbogognon winterianus Jowitt (citronella) EOs. We have shown that the application of the whole EOs and their major individual compounds on the leaves and cotyledons of A. thaliana appears to be promising: when applied on cotyledons or leaves, EOs induce damages that are as important as those observed for commercial herbicides. Since EOs are small amphiphilic molecules, they can cross the mesh of cell wall and interact directly with the plant plasma membrane (PPM). Modifying the lipid organization could lead to crucial cellular effects, notably on protein function. We used a unique and original combination of in silico (molecular dynamics simulations) and in vitro (Langmuir monolayers, isothermal calorimetry, fluorescence and infrared spectroscopies) biophysical approaches, previously developed to study structure-function relationships of molecules of biological interest (pharmacological drugs , proteins, peptides, surfactants…) to investigate the interaction of EOs or their individual compounds with bio-mimetic plant plasma membranes to better understand the structure- activity relationships in the context of their bioherbicide activity. [less ▲]

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See detailCynara cardunculus crude extract as a powerful natural herbicide and insight into the mode of action of its bioactive molecules
Ben Kaab, Sofiène ULiege; Lins, Laurence ULiege; Hanafi, Marwa ULiege et al

in Biomolecules (2020), 10(2), 209

The use of chemical herbicides could not only potentially induce negative impacts on the environment, animals, and human health, but also increase the weed resistance to herbicides. In this context, the ... [more ▼]

The use of chemical herbicides could not only potentially induce negative impacts on the environment, animals, and human health, but also increase the weed resistance to herbicides. In this context, the use of plant extracts could be an interesting and natural alternative to chemical products. It is important to understand the mode of action of their bioactive compounds. This is why we have studied the herbicidal effect of Cynara cardunculus crude extract in terms of inhibition of weeds’ seedling growth and its impact on physiological parameters of treated plantlets, like conductivity, dry weight, and fluorescence, and biochemical parameters linked to oxidative stress. We have observed that C. cardunculus crude extract induces oxidative stress in the treated plants and consequently disturbs the physiological and biochemical functions of the plant cells. We have investigated the herbicidal activity of three bioactive compounds, naringenin, myricitrin, and quercetin, from the C. cardunculus crude extract. In both pre- and post-emergence trials, naringenin and myricitrin were significantly more phytotoxic than quercetin. We suggest that their differential initial interaction with the plant’s plasma membrane could be one of the main signals for electrolyte leakage and production of high levels of phenoxyl radicals. [less ▲]

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See detailThe Trypanosoma Brucei KIFC1 Kinesin Ensures the Fast Antibody Clearance Required for Parasite Infectivity.
Lecordier, Laurence; Uzureau, Sophie; Vanwalleghem, Gilles et al

in iScience (2020), 23(9), 101476

Human innate immunity to Trypanosoma brucei involves the trypanosome C-terminal kinesin TbKIFC1, which transports internalized trypanolytic factor apolipoprotein L1 (APOL1) within the parasite. We show ... [more ▼]

Human innate immunity to Trypanosoma brucei involves the trypanosome C-terminal kinesin TbKIFC1, which transports internalized trypanolytic factor apolipoprotein L1 (APOL1) within the parasite. We show that TbKIFC1 preferentially associates with cholesterol-containing membranes and is indispensable for mammalian infectivity. Knockdown of TbKIFC1 did not affect trypanosome growth in vitro but rendered the parasites unable to infect mice unless antibody synthesis was compromised. Surface clearance of Variant Surface Glycoprotein (VSG)-antibody complexes was far slower in these cells, which were more susceptible to capture by macrophages. This phenotype was not due to defects in VSG expression or trafficking but to decreased VSG mobility in a less fluid, stiffer surface membrane. This change can be attributed to increased cholesterol level in the surface membrane in TbKIFC1 knockdown cells. Clearance of surface-bound antibodies by T. brucei is therefore essential for infectivity and depends on high membrane fluidity maintained by the cholesterol-trafficking activity of TbKIFC1. [less ▲]

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See detailStress-related accumulation of arabidopsides: impact on plant chloroplasts - Flash presentation
Genva, Manon ULiege; Deleu, Magali ULiege; Andersson, Mats X et al

Conference (2020)

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See detailAmphiphilic azobenzenes: Antibacterial activities and biophysical investigation of their interaction with bacterial membrane lipids
Franche, Antoine; Fayeulle, Antoine; Lins, Laurence ULiege et al

in Bioorganic Chemistry (2020), 94

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See detailStress-related accumulation of arabidopsides: impact on plant chloroplasts
Genva, Manon ULiege; Deleu, Magali ULiege; Andersson, Mats X et al

Poster (2020)

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See detailEnhancing the Membranolytic Activity of Chenopodium quinoa Saponins by Fast Microwave Hydrolysis
Colson, Emmanuel; Savarino, Philippe; Claereboudt, Emily ULiege et al

in Molecules (2020), 25(7), 1731

Saponins are plant secondary metabolites. There are associated with defensive roles due to their cytotoxicity and are active against microorganisms. Saponins are frequently targeted to develop efficient ... [more ▼]

Saponins are plant secondary metabolites. There are associated with defensive roles due to their cytotoxicity and are active against microorganisms. Saponins are frequently targeted to develop efficient drugs. Plant biomass containing saponins deserves sustained interest to develop high-added value applications. A key issue when considering the use of saponins for human healthcare is their toxicity that must be modulated before envisaging any biomedical application. This can only go through understanding the saponin-membrane interactions. Quinoa is abundantly consumed worldwide, but the quinoa husk is discarded due to its astringent taste associated with its saponin content. Here, we focus on the saponins of the quinoa husk extract (QE). We qualitatively and quantitively characterized the QE saponins using mass spectrometry. They are bidesmosidic molecules, with two oligosaccharidic chains appended on the aglycone with two different linkages; a glycosidic bond and an ester function. The latter can be hydrolyzed to prepare monodesmosidic molecules. The microwave-assisted hydrolysis reaction was optimized to produce monodesmosidic saponins. The membranolytic activity of the saponins was assayed based on their hemolytic activity that was shown to be drastically increased upon hydrolysis. In silico investigations confirmed that the monodesmosidic saponins interact preferentially with a model phospholipid bilayer, explaining the measured increased hemolytic activity. [less ▲]

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See detailContributions and Limitations of Biophysical Approaches to Study of the Interactions between Amphiphilic Molecules and the Plant Plasma Membrane
Furlan, Aurélien ULiege; Laurin, Yoann ULiege; Botcazon, Camille et al

in Plants (2020), 9(5), 648

Some amphiphilic molecules are able to interact with the lipid matrix of plant plasma membranes and trigger the immune response in plants. This original mode of perception is not yet fully understood and ... [more ▼]

Some amphiphilic molecules are able to interact with the lipid matrix of plant plasma membranes and trigger the immune response in plants. This original mode of perception is not yet fully understood and biophysical approaches could help to obtain molecular insights. In this review, we focus on such membrane-interacting molecules, and present biophysically grounded methods that are used and are particularly interesting in the investigation of this mode of perception. Rather than going into overly technical details, the aim of this review was to provide to readers with a plant biochemistry background a good overview of how biophysics can help to study molecular interactions between bioactive amphiphilic molecules and plant lipid membranes. In particular, we present the biomimetic membrane models typically used, solid-state nuclear magnetic resonance, molecular modeling, and fluorescence approaches, because they are especially suitable for this field of research. For each technique, we provide a brief description, a few case studies, and the inherent limitations, so non-specialists can gain a good grasp on how they could extend their toolbox and/or could apply new techniques to study amphiphilic bioactive compound and lipid interactions [less ▲]

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See detailPlant-pathogen interactions: underestimated roles of phyto-oxylipins
Deboever, Estelle ULiege; Deleu, Magali ULiege; Mongrand, Sébastien et al

in Trends in Plant Science (2020), 25(1), 22-34

Plant oxylipins are produced under a wide range of stress conditions, and even though they are well known to activate stress-related signalling pathways, the non-signalling roles of phyto-oxylipins are ... [more ▼]

Plant oxylipins are produced under a wide range of stress conditions, and even though they are well known to activate stress-related signalling pathways, the non-signalling roles of phyto-oxylipins are poorly understood. We describe oxylipins as direct biocidal agents and propose that structure-function relationships play here a pivotal role. Indeed, based on their chemical configuration, plant oxylipins, such as reactive oxygen and electrophile species, activate defence-related genes expression. We also propose that their ability to interact with pathogen membranes is important, but still misunderstood, and that they are involved in cross-kingdom communication. Taken as a whole, the current literature suggests that plant oxylipins have a high potential as biocontrol agents. However, the mechanisms underlying these multi-faceted compounds remain largely unknown. [less ▲]

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See detail« SAR out of the box » : insights into the relations between molecular structure, membrane interaction and activity of biomolecules using complementary biophysical tools
Lins, Laurence ULiege; Deleu, Magali ULiege

Conference (2019, December 02)

Classically, structure-Activity Relationship (SAR) is an approach designed to find relationships between chemical structure (or structural-related properties), 3D structure and biological activity of ... [more ▼]

Classically, structure-Activity Relationship (SAR) is an approach designed to find relationships between chemical structure (or structural-related properties), 3D structure and biological activity of compounds of interest, as defined by Crum-Brown and Fraser in 1865. This is because similar compounds should have similar physico-chemical and biological properties. If this notion is almost always associated to pharmacological drug design and assessment of side effects of existing compounds in the human health field, it can actually be extended to a large number of bioactive molecules exerting a number of cellular activities. For instance, interactions between plant or bacterial secondary metabolites as well as proteins belonging to the same structural family could have differential activities, notably at the level of the cellular membrane. In this talk, I propose to overview some complementary “in vitro” and “in silico” biophysical approaches such as those described in ref [1–3] that can give information about the relation between chemical structure (or structural-related properties), 3D structure and membrane activity of bioactive molecules. [less ▲]

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