A5059726973- Lipid Membrane Structure and Behavior
- Sphingolipid Metabolism and Signaling
- Cellular transport and secretion
- Erythrocyte Function and Pathophysiology
- Lysosomal Storage Disorders Research
- RNA Interference and Gene Delivery
- DNA and Nucleic Acid Chemistry
- Surfactants and Colloidal Systems
- Metabolomics and Mass Spectrometry Studies
- Nanoparticle-Based Drug Delivery
- Lipid metabolism and biosynthesis
- Porphyrin and Phthalocyanine Chemistry
- Nanopore and Nanochannel Transport Studies
- Force Microscopy Techniques and Applications
- Peroxisome Proliferator-Activated Receptors
- Inhalation and Respiratory Drug Delivery
- Neonatal Health and Biochemistry
- Photoreceptor and optogenetics research
- Advanced biosensing and bioanalysis techniques
- Photochromic and Fluorescence Chemistry
- Hydrogels: synthesis, properties, applications
- Pancreatitis Pathology and Treatment
- Biomedical Research and Pathophysiology
- Mass Spectrometry Techniques and Applications
- RNA and protein synthesis mechanisms
University of the Basque Country
2010-2024
Instituto Biofisika
2018-2024
University of Geneva
2019-2023
NCCR Chemical Biology - Visualisation and Control of Biological Processes Using Chemistry
2019-2023
Fundación Biofísica Bizkaia
2018-2023
Ikerbasque
2023
Tools to image membrane tension in response mechanical stimuli are badly needed mechanobiology. We have recently introduced mechanosensitive flipper probes report quantitatively global changes fluorescence lifetime imaging microscopy (FLIM) images of living cells. However, address specific questions on physical forces biology, the need be localized precisely interest (MOI). Herein we present a general strategy MOI by tagging our newly HaloFlippers self-labeling HaloTags fused proteins this...
To image membrane tension in selected membranes of interest (MOI) inside living systems, the field mechanobiology requires increasingly elaborated small-molecule chemical tools. We have recently introduced HaloFlipper, i.e., a mechanosensitive flipper probe that can localize MOI using HaloTag technology to report local changes fluorescence lifetime imaging microscopy. However, linker tethering hampers lateral diffusion all lipid domains MOI. For more global measurement any MOI, we present...
The mechanisms of Pb(II) toxicity have been studied in human red blood cells using confocal microscopy, immunolabeling, fluorescence-activated cell sorting and atomic force microscopy. process follows a sequence events, starting with calcium entry, followed by potassium release, morphological change, generation ceramide, lipid flip-flop finally lysis. Clotrimazole blocks channels the whole is inhibited. Immunolabeling reveals ceramide-enriched domains linked to while use neutral...
Abstract Giant unilamellar vesicles (GUVs) are model cell-sized systems that have broad applications including drug delivery, analysis of membrane biophysics, and synthetic reconstitution cellular machineries. Although numerous methods for the generation free-floating GUVs been established over past few decades, only a fraction successfully produced uniform vesicle populations both from charged lipids in buffers physiological ionic strength. In method described here, we generate large...
Abstract The lipid composition of cellular membranes is dynamic and undergoes remodelling affecting biophysical properties, such as membrane fluidity, which are critical to biological function. Here, we introduce an optical approach manipulate fluidity based on exogenous synthetic fatty acid with azobenzene photoswitch, termed FAAzo4 . Cells rapidly incorporate into phosphatidylcholine (PC), the major phospholipid in mammalian cells, a concentration- cell type-dependent manner. This...
This article describes four fluorescent membrane tension probes that have been designed, synthesized, evaluated, commercialized and applied to current biology challenges in the context of NCCR Chemical Biology. Their names are Flipper-TR®, ER Lyso Mito Flipper-TR®. They available from Spirochrome.
This study was aimed at preparing and characterizing plasma membranes (PM) from Chinese Hamster Ovary (CHO) cells. Two methods of PM preparation were applied, one based on adhering cells to a poly-lysine-coated surface, followed by hypotonic lysis removal intracellular components, so that patches remain adhered each other, second consisting bleb induction in cells, separation giant membrane vesicles (GPMV). Both gave rise sufficient amounts allow biophysical biochemical characterization....
Sphingolipids (SL) are ubiquitous in mammalian cell membranes, yet there is little data on the behavior of cells under SL-restriction conditions. LY-B derive from a CHO linein whichserine palmitoyl transferase (SPT), thus de novo SL synthesis, suppressed, while maintaining capacity taking up and metabolizing exogenous sphingoid bases culture medium. In this study, were adapted to grow fetal bovine serum (FBS)-deficient medium avoid external uptake lipids. The lowest FBS concentration that...
Abstract Suppression of a specific gene effect can be achieved by genetic as well chemical methods. Each approach may hide unexpected drawbacks, usually in the form side effects. In present study, inhibitor myriocin was used to block serine palmitoyltransferase (SPT), first enzyme sphingolipid synthetic pathway, CHO cells. The subsequent biophysical changes plasma membranes were measured and compared with results obtained genetically modified cell line containing defective SPT (the LY-B...
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The discovery of a novel sphingolipid subclass, the (1-deoxy)sphingolipids, which lack 1-hydroxy group, attracted considerable attention in last decade, mainly due to their involvement disease. They differed physico-chemical properties from canonical (or 1-hydroxy) sphingolipids and they were more toxic when accumulated cells, inducing neurodegeneration other dysfunctions. (1-Deoxy)ceramides, (1-deoxy)dihydroceramides, (1- deoxymethyl)dihydroceramides, latter two containing saturated...
ABSTRACT Sphingolipids have been shown to play important roles in physiology and cell biology, but a systematic examination of their functions is lacking. We performed genome-wide CRISPRi screen sphingolipid-depleted cells identified hypersensitive mutants genes membrane trafficking lipid biosynthesis, including ether synthesis. Systematic lipidomic analysis showed coordinate regulation lipids with sphingolipids, where depletion one these types resulted increases the other, suggesting an...
ABSTRACT Sphingolipids (SL) are ubiquitous in mammalian cell membranes, yet there is little data on the behavior of cells under SL-restriction conditions. LY-B derive from a CHO line which serine palmitoyl transferase (SPT), thus de novo SL synthesis, suppressed, while maintaining capacity taking up and metabolizing exogenous sphingoid bases culture medium. In present study were adapted to grow fetal bovine serum (FBS)-deficient medium avoid external uptake lipids. The lowest FBS...