Dynamique des lipides cellulaires

Durant ces dernières années, la notion de micro- domaines à la surface des membranes cellulaires a vu le jour. Il s’agit de « lipid rafts » qui impliquent des sphingoglycolipides (SGL) spécifiques et du cholestérol, auxquels sont associées des protéines particulières qui comportent, soit un ancrage lipidique, soit des propriétés qui en font des constituants transmembranaires. Ces complexes ciblent ces protéines à la surface des membranes cellulaires, et les composés lipidiques leur restent en général associés. Ces « rafts » sont également impliqués dans la constitution de récepteurs de surface et dans les phénomènes d’endocytose qui suivent l’action de leurs ligands. Ces « rafts » constituent des structures très compactes, insolubles dans les détergents non ionisés à basse température, ce qui permet leur isolement en tant que complexes. De nombreux « rafts » différents peuvent être isolés en fonction des détergents utilisés; leur multiplicité peut être liée à l’hétérogénéité des SGL qui pourrait être nécessaire pour le ciblage de protéines spécifiques. Les SGL ont une grande complexité et diversité selon les organes, les tissus, les types cellulaires. Ils sont impliqués dans les phénomènes de polarisation cellulaire. Ils sont modifiés au cours de la différenciation cellulaire, du cancer. Ils peuvent être l’objet d’anticorps dans les neuropathies dysimmunitaires, et impliqués dans des maladies dégénératives du système nerveux, d’origine génétique (neurolipidoses). Les altérations du cholestérol telles qu’on les observe dans la maladie de Niemann Pick de type C et dans la maladie d’Alzheimer ont aussi un retentissement sur le fonctionnement du système nerveux. Ainsi ces microdomaines spécialisés sont impliqués dans la dynamique cellulaire normale et pathologique

Mechanisms governing the fragmentation of glycerophospholipids containing choline and ethanolamine polar head groups

International audienceGlycerophospholipids are the major amphiphilic molecules found in the plasma membrane bilayer of all vertebrate cells. Involved in many biological processes, their huge structural diversity and large concentration scale make their thorough characterization extremely difficult in complex biological matrices. Mass spectrometry techniques are now recognized as being among the most powerful methods for the sensitive and comprehensive characterization of lipids. Depending on the experimental conditions used during electrospray ionization mass spectrometry experiments, glycerophospholipids can be detected as different molecular species (e.g. protonated, sodiated species) when analyzed either in positive or negative ionization modes or by direct introduction or hyphenated mass spectrometry-based methods. The observed ionized forms are characteristic of the corresponding phospholipid structures, and their formation is highly influenced by the polar head group. Although the fragmentation behavior of each phospholipid class has already been widely studied under low collision energy, there are no established rules based on charge-induced dissociation mechanisms for explaining the generation of fragment ions. In the present paper, we emphasize the crucial roles played by ion-dipole complexes and salt bridges within charge-induced dissociation processes. Under these conditions, we were able to readily explain almost all the fragment ions obtained under low-energy collision-induced dissociation for particular glycerophospholipids and lysoglycerophospholipids species including glycerophosphatidylcholines and glycerophosphatidylethanolamines. Thus, in addition to providing a basis for a better comprehension of phospholipid fragmentation processes, our work also highlighted some potentially new relevant diagnostic ions to signal the presence of particular lipid species

Multicentric investigations of the role in the disease severity of accelerated phospholipid changes in COVID-19 patient airway

International audienceContext. The changes in host membrane phospholipids are crucial in airway infection pathogenesis. Phospholipase A2 hydrolyzes host cell membranes, producing lyso-phospholipids and free fatty acids, including arachidonic acid (AA), which contributes significantly to lung inflammation. Aim. Follow these changes and their evolution from day 1, day 3 to day 7 in airway aspirates of 89 patients with COVID-19-associated acute respiratory distress syndrome and examine whether they correlate with the severity of the disease. The patients were recruited in three French intensive care units. The analysis was conducted from admission to the intensive care unit until the end of the first week of mechanical ventilation. Results. In the airway aspirates, we found significant increases in the levels of host cell phospholipids, including phosphatidyl-serine and phosphatidyl-ethanolamine, and their corresponding lyso-phospholipids. This was accompanied by increased levels of AA and its inflammatory metabolite prostaglandin E2 (PGE2). Additionally, enhanced levels of ceramides, sphingomyelin, and free cholesterol were observed in these aspirates. These lipids are known to be involved in cell death and/or apoptosis, whereas free cholesterol plays a role in virus entry and replication in host cells. However, there were no significant changes in the levels of dipalmitoyl-phosphatidylcholine, the major surfactant phospholipid. A correlation analysis revealed an association between mortality risk and levels of AA and PGE2, as well as host cell phospholipids. Conclusion. Our findings indicate a correlation between heightened cellular phospholipid modifications and variations in AA and PGE2 with the severity of the disease in patients. Nevertheless, there is no indication of surfactant alteration in the initial phases of the illness

Mechanistic study of competitive releases of H2O, NH3 and CO2 from deprotonated aspartic and glutamic acids: Role of conformation

The aims of this study were to highlight the impact of minor structural differences (e.g. an aminoacid side chain enlargement by one methylene group), on ion dissociation under collision-induced dissociation conditions, and to determine the underlying chemical mechanisms. Therefore, we compared fragmentations of deprotonated aspartic and glutamic acids generated in negative electrospray ionization. Energy-resolved mass spectrometry breakdown curves were recorded and MS3 experiments performed on an Orbitrap Fusion for high-resolution and high-mass accuracy measurements. Activated fragmentations were performed using both the resonant and non-resonant excitation modes (i.e., CID and HCD, respectively) in order to get complementary information on the competitive and consecutive dissociative pathways. These experiments showed a specific loss of ammonia from the activated aspartate but not from the activated glutamate. We mainly focused on this specific observed loss from aspartate. Two different mechanisms based on intramolecular reactions (similar to those occurring in organic chemistry) were proposed, such as intramolecular elimination (i.e. Ei-like) and nucleophilic substitution (i.e. SNi-like) reactions, respectively, yielding anions as fumarate and α lactone from a particular conformation with the lowest steric hindrance (i.e. with antiperiplanar carboxyl groups). The detected deaminated aspartate anion can then release CO2 as observed in the MS3 experimental spectra. However, quantum calculations did not indicate the formation of such a deaminated aspartate product ion without loss of carbon dioxide. Actually, calculations displayed the double neutral (NH3+CO2) loss as a concomitant pathway (from a particular conformation) with relative high activation energy instead of a consecutive process. This disagreement is apparent since the concomitant pathway may be changed into consecutive dissociations according to the collision energy i.e., at higher collision energy and at lower excitation conditions, respectively. The latter takes place by stabilization of the deaminated aspartate solvated with two residual molecules of water (present in the collision cell). This desolvated anion formed is an α lactone substituted by a methylene carboxylate group. The vibrational excitation acquired by [(D−H)−NH3]−during its isolation is enough to allow its prompt decarboxylation with a barrier lower than 8.4 kJ/mol. In addition, study of glutamic acid-like diastereomers constituted by a cyclopropane, hindering any side chain rotation, confirms the impact of the three-dimensional geometry on fragmentation pathways. A significant specific loss of water is only observed for one of these diastereomers. Other experiments, such as stable isotope labeling, need to be performed to elucidate all the observed losses from activated aspartate and glutamate anions. These first mechanistic interpretations enhance understanding of this dissociative pathway and underline the necessity of studying fragmentation of a large number of various compounds to implement properly new algorithms for de novo elucidation of unknown metabolites

Metabolomic profiling of cardiac allografts after controlled circulatory death

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