Lateral stress profile and fluorescent lipid probes. FRET pair of probes that introduces minimal distortions into lipid packing

Tretiakova, Daria; Алексеева, А. С.; Galimzyanov, Timur R.; Boldyrev, A.M.; Chernyadyev, Andrey Yu.; Ermakov, Yu. A.; Batishchev, Oleg V.; Водовозова, Е. Л.; Болдырев, Иван

doi:10.1016/j.bbamem.2018.05.020

Cited by 22 publications

(19 citation statements)

References 35 publications

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“…First, the interaction between liposomes and albumin was examined using Förster resonance energy transfer (FRET) using BSA-FITC conjugate as a donor and BCHB-PC probe in liposome bilayer as an acceptor molecule [45]. Provided albumin adsorbed on a liposome surface, FITC signal should decrease due to the energy transfer to BCHB-PC proportionally to the amount of the protein adsorbed, depth of its penetration in the bilayer, and/or changes of mutual arrangement of the fluorophores.…”

Section: Characteristics Of the Liposomesmentioning

confidence: 99%

Phosphatidylinositol Stabilizes Fluid-Phase Liposomes Loaded with a Melphalan Lipophilic Prodrug

et al. 2021

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Previously, a liposomal formulation of a chemotherapeutic agent melphalan (Mlph) incorporated in a fluid lipid bilayer of natural phospholipids in the form of dioleoylglyceride ester (MlphDG) was developed and the antitumor effect was confirmed in mouse models. The formulation composed of egg phosphatidylcholine (ePC), soybean phosphatidylinositol (PI), and MlphDG (8:1:1, by mol) showed stability in human serum for at least 4–5 h. On the contrary, replacing PI with pegylation of the liposomes, promoted fast dissociation of the components from the bilayer. In this work, interactions of MlphDG-liposomes with the most abundant plasma protein—albumin—in function of the presence of PI in the formulation were explored using Fourier transform infrared spectroscopy. The release of MlphDG from the liposomes was studied by asymmetrical flow field-flow fractionation (AF4) using micelles formed by a polyethylene glycol conjugate with phosphatidylethanolamine to mimic the physiological lipid sink like lipoproteins. Our results show that PI actually protects the membrane of MlphDG-liposomes from the protein penetration, presumably due to pairing between the positively charged MlphDG and negatively charged PI, which compensates for the heterogeneity of the lipid bilayer. The AF4 technique also evidences high stability of the formulation as a drug carrier.

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Section: Characteristics Of the Liposomesmentioning

confidence: 99%

Phosphatidylinositol Stabilizes Fluid-Phase Liposomes Loaded with a Melphalan Lipophilic Prodrug

et al. 2021

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“…TMB-PC were obtained. When the ratio of probe molecules to lipids is 1/4000, no energy transfer is observed between neighboring TMB fluorophores, and the fluorescence anisotropy depends only on the fluorophore mobility (see also [17,18]).…”

Section: Methodsmentioning

confidence: 99%

“…In membranes, the fluorescence anisotropy of the label depends on the packing density of lipids and their mobility. The maximum value of TMB anisotropy (in propylene glycol solution at -70°C) is 0.39, which corresponds to a completely inhibited probe [18]. Changes in the anisotropy of TMB-PC fluorescence in the lipid membrane ranges from 0.18 (gel phase) to 0.10 (liquid crystal phase) [18].…”

Section: Registration Of Changes In the Membranementioning

confidence: 97%

“…Monitoring the change in the state of the membrane is carried out using fluorescent lipid probes (by fluorescence anisotropy). In this case, TMB-PC (BODIPY-labeled phosphatidylcholine) is used [18], but other membrane probes can also be used. For example, for laurdan and prodan, it may be more convenient to record changes in the emission spectrum…”

Section: Registration Of Changes In the Membranementioning

confidence: 99%

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Estimation of the Phospholipase A2 Selectivity on POPC/POPG Membranes Using the Interaction Map

Алексеева

Volynsky

Болдырев

2021

Biochem. Moscow Suppl. Ser. A

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The regulation of the activity and selectivity of phospholipase A2 (PLA2), which is capable of cleaving fatty acid in the second position (sn-2) of the phospholipid, is carried out through the membrane-binding and catalytic sites of the enzyme. For hydrolytic activity, PLA2 must first bind to the phospholipid membrane, and the binding efficiency depends on the composition of the membrane. The membrane-binding site of PLA2 is formed by several tens of amino acids and its composition differs from enzyme to enzyme; hydrophobic and positively charged amino acids play a key role in the interaction. In this work, we investigated the interaction of PLA2 from bee venom with phospholipid bilayers of palmitoyl oleoylphosphatidylcholine (POPC) containing different amounts of palmitoyloleoylphosphatidylglycerol (POPG). On the basis of the measurements of the protein intrinsic fluorescence and the anisotropy of the fluorescence of the lipid probe we propose the construction of lipid–protein interaction maps, which reflect both the efficiency of protein binding and changes in the structure of the membrane. These changes cause alterations in the fluorescence anisotropy of the label, which in turn is a measure of the mobility of the lipid environment of the fluorescent probe. Analysis of interaction maps showed that there is a relationship between lipid mobility and enzyme binding efficiency: the optimum interaction of PLA2 with membranes from a POPC/POPG mixture lies in the region of the highest lipid mobility, and not in the region of the highest negative charge. This dependence complements the existing understanding of the process of recognition of the membrane surface by the enzyme and the selection of lipids by the enzyme already bound to the membrane. The proposed mapping method can be extended to other membrane-active proteins.

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“…Despite the value of ratiometric FRET, one of its main limitations is its dependence on relative quantifications rather than absolute measurements. So far, FRET has not been reported in vivo in parasitology, yet could be very useful to monitor the activity of kinases, proteases, GTPases [24]; calcium changes [25]; lipid concentrations [23,[26][27][28] during infection; and even physical stresses such as temperature, mechanical stressors, or electromagnetic fields [29], as well as molecular interactions between host and pathogens; host cell subsets, or parasite molecules. Likewise, there has been significant progress in the development of software to allow separate quantification of signals from single cells in complex 3D environments, such as those present in organs of living animals [30,31].…”

mentioning

confidence: 99%

Toolbox for In Vivo Imaging of Host–Parasite Interactions at Multiple Scales

Niz

Spadin

Marti

et al. 2019

Trends in Parasitology

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Animal models have for long been pivotal for parasitology research. Over the last few years, techniques such as intravital, optoacoustic and magnetic resonance imaging, optical projection tomography, and selective plane illumination microscopy developed promising potential for gaining insights into host-pathogen interactions by allowing different visualization forms in vivo and ex vivo. Advances including increased resolution, penetration depth, and acquisition speed, together with more complex image analysis methods facilitate tackling biological problems previously impossible to study and/or quantify. Here we discuss advances and challenges in the in vivo imaging toolbox, which hold important potential for the field of parasitology. KeywordsParasitology, imaging, in vivo, animal models Imaging toolbox in parasitologyImaging techniques developed for biomedical applications have had an important impact in parasitology research. Such techniques include platforms developed to image host-pathogen interactions at various scales, ranging from molecules to whole organisms (summarised in table 1). These techniques have complementary advantages with respect to each other. This review focuses on the technological advances used for visualization of host-pathogen interactions either in vivo, or ex vivo in whole organisms in five imaging techniques: intravital microscopy (IVM), optical projection tomography (OPT), bioluminescence imaging, optoacoustic imaging (OAI), and magnetic resonance imaging (MRI). Advanced fluorescence methods applied to intravital microscopyIntravital microscopy (IVM) is a powerful technique to investigate dynamic cellular processes and host-parasite interactions within functioning organs. Organs studied by IVM in the context of parasitology include the brain [1][2][3][4], the skin [5][6][7], the placenta [8,9], the lungs [10], the liver [11][12][13], and the spleen [14,15] (summarized in table 2, green=IVM exists; yellow=organ relevant but IVM never done; grey = IVM not done). Important advances in parasitology have been achieved using wide-field epifluorescence, confocal, spinning disc, or two-photon IVM.Recent developments, which have expanded the applications of IVM include the generation of

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Lateral stress profile and fluorescent lipid probes. FRET pair of probes that introduces minimal distortions into lipid packing

Cited by 22 publications

References 35 publications

Phosphatidylinositol Stabilizes Fluid-Phase Liposomes Loaded with a Melphalan Lipophilic Prodrug

Phosphatidylinositol Stabilizes Fluid-Phase Liposomes Loaded with a Melphalan Lipophilic Prodrug

Estimation of the Phospholipase A2 Selectivity on POPC/POPG Membranes Using the Interaction Map

Toolbox for In Vivo Imaging of Host–Parasite Interactions at Multiple Scales

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