Université PSL

Publications

RECHERCHER

Laboratoire :
Auteur :
Revue :
Année :

A L'ATTENTION DES EQUIPES IPGG :

- Pour toute publication de résultats ayant reçu l’aide de l’IPGG (présence dans les locaux de l’IPGG, passage sur la plateforme technologique de l’IPGG, collaboration inter équipes IPGG, lié à une bourse doctorale ou postdoctorale IPGG, ou encore utilisation des espaces communs), il vous faut indiquer  cette phrase « Ce travail a été réalisé avec le soutien du laboratoire d’excellence Institut Pierre-Gilles de Gennes (programme Investissements d’avenir ANR-10-IDEX-0001-02 PSL et ANR-10-LABX-31). » / « This work has received the support of "Institut Pierre-Gilles de Gennes" (laboratoire d’excellence, “Investissements d’avenir” program ANR-10-IDEX-0001-02 PSL and ANR-10-LABX-31.). ».

- Pour toute publication de résultats obtenu via l'utilisation d’un équipement acheté par l’Equipex IPGG, il vous faut ajouter  la codification suivante : « ANR-10-EQPX-34 ».

An Epigenetic Priming Mechanism Mediated by Nutrient Sensing Regulates Transcriptional Output
Laboratoire pour la biologie quantitative du développement - Natalia Stec, Katja Doerfel, Kelly Hills-Muckey, Victoria M. Ettorre, Sevinc Ercan, Wolfgang Keil, C. M. Hammell
bioRxiv - - doi.org/10.1101/2020.09.01.278127 - 2020
While precise tuning of gene expression levels is critical for most developmental pathways, the mechanisms by which the transcriptional output of dosage-sensitive molecules is established or modulated by the environment remain poorly understood. Here, we provide a mechanistic framework for how the conserved transcription factor BLMP-1/Blimp1 operates as a pioneer factor to decompact chromatin near its target loci hours before transcriptional activation and by doing so, regulates both the duration and amplitude of subsequent target gene transcription. This priming mechanism is genetically separable from the mechanisms that establish the timing of transcriptional induction and functions to canalize aspects of cell-fate specification, animal size regulation, and molting. A key feature of the BLMP-1-dependent transcriptional priming mechanism is that chromatin decompaction is initially established during embryogenesis and maintained throughout larval development by nutrient sensing. This anticipatory mechanism integrates transcriptional output with environmental conditions and is essential for resuming normal temporal patterning after animals exit nutrient-mediated developmental arrests.
Electrocatalytic behaviour of CeZrOx-supported Ni catalysts in plasma assisted CO2 methanation
Laboratoire Procédés - Plasmas - Microsystèmes - Maria Mikhail, Patrick Da Costa, Jacques Amouroux, Siméon Cavadias, Michael Tatoulian, Stéphanie Ognier and María Elena Gálvez
Catalys Science & Technology - 10 4532-4543 - https://doi.org/10.1039/D0CY00312C - 2020
Plasma-catalytic and thermo-catalytic methanation were assayed in the presence of a CeZrOx-supported Ni catalyst, proving that high CO2 conversions and high methane yields can be obtained under dielectric barrier discharge (DBD) plasma conditions and that they are maintained with time-on-stream over 100 h operating time. The characterization of the spent catalysts through TPD-MS, ATR-FTIR, TEM and HR-TEM and XRD evidenced the coexistence of a Ni0/NiO phase together with an increased presence of Ce3+ cations and oxygen vacancies, on the surface of the catalyst submitted to plasma catalytic operation. The different facts collected through physicochemical characterization point to our catalyst behaving like a PN junction, or like a fuel cell, with a P-side, the anode, i.e. the Ni-side releasing electrodes, while the CeZrOx support, N-side and cathode, acts as an acceptor. The DBD plasma, rich in ionic species and free electrodes, acts as the electrolyte, conducting the electrodes in the right direction. Oxygen accumulation on the surface of the catalyst during thermo-catalytic methanation leads to the formation of non-reactive adsorbed species, whereas Ni-sintering is favored. Under DBD plasma conditions, electron transfer is guaranteed and the adsorption–desorption of reactants and products is favored.
Coupling experiment and simulation analysis to investigate physical parameters of CO2 methanation in a plasma catalytic hybrid process
Laboratoire Procédés - Plasmas - Microsystèmes - Bo Wang Maria Mikhail Maria Elena Galvez Simeon Cavadias Michael Tatoulian Patrick Da Costa Stéphanie Ognier
First published - 17 9 - https://doi.org/10.1002/ppap.201900261 - 2020
This study focuses on the use of a heterogeneous catalyst Ni/Ce0.58Zr0.42O2 to study the Sabatier reaction in conventional catalytic thermal heating and the dielectric barrier discharge plasma‐catalytic process. Its aim is to study the threshold temperature of the Sabatier reaction in plasma conditions. A set of experiments with different inlet flow rates is carried out in a plasma reactor to investigate the steady‐state temperature of the reaction. To estimate the threshold temperature of the Sabatier reaction more accurately, the temperature difference between the catalytic bed and the external surface of the reactor is calculated and simulated in COMSOL Multiphysics® software. Finally, the threshold temperature of the Sabatier reaction during plasma processing is assumed to be 116°C, based on the experimental data and simulation analysis.
Ni-Fe layered double hydroxide derived catalysts for non-plasma and DBD plasma-assisted CO2 methanation
Laboratoire Procédés - Plasmas - Microsystèmes - D Moreno, MV Ognier, S Motak, Grzybek, T Da Costa, P Galvez
Catalys Science & Technology - 45 17 - DOI: 10.1016/j.ijhydene.2019.06.095 - 2020
A series of bi-metallic layered double hydroxide derived materials, containing a fixed amount of Ni promoted with various amounts of Fe were obtained by co-precipitation. The synthesized materials were characterized by X-ray diffraction (XRD), temperature-programmed reduction (H 2-TPR), temperature-programmed desorption of CO 2 (CO 2-TPD), elemental analysis and low temperature N 2 sorption and tested as catalysts in CO 2 methanation at atmospheric pressure. The obtained results confirmed the formation of mixed nano-oxides after thermal decomposition of the precursor and suggest successful introduction of both nickel and iron into the layers of Layered Double Hydroxides (LDHs). The introduction of Fe into the layered double hydroxides changed the interaction between Ni and supports matrix as proven by temperature programmed reduction (H 2-TPR). The introduction of low amount of iron influenced positively the catalytic activity in CO 2 methanation at 250 C, with CO 2 conversion increasing from 21% to 72% with CH 4 selec-tivity ranging from 97 to 99% at 250 C. No other products, except CH 4 and CO were registered during the experiments. In order to enhance the catalytic activity a non-thermal plasma created by dielectric barrier discharge was applied. The obtained results prove that * Corresponding author.
Plasma-Induced Polymerizations: A New Synthetic Entry in Liquid Crystal Elastomer Actuators
Laboratoire Procédés - Plasmas - Microsystèmes - Bin Ni Mengxue Zhang Cédric Guyon Patrick Keller Michael Tatoulian Min
First published - 41 19 - https://doi.org/10.1002/marc.202000385 - 2020
The research on soft actuators including liquid crystal elastomers (LCEs) becomes more and more appealing at a time when the expansion of artificial systems is blooming. Among the various LCE actuators, the bending deformation is often in the origin of many actuation modes. Here, a new strategy with plasma technology is developed to prepare single‐layer main‐chain LCEs with thermally actuated bending and contraction deformations. Two distinct reactions, plasma polymerization and plasma‐induced photopolymerization, are used to polymerize in one step the nematic monomer mixture aligned by magnetic field. The plasma polymerization forms cross‐linked but disoriented structures at the surface of the LCE film, while the plasma‐induced photopolymerization produces aligned LCE structure in the bulk. The actuation behaviors (bending and/or contraction) of LCE films can be adjusted by plasma power, reaction time, and sample thickness. Soft robots like crawling walker and flower mimic are built by LCE films with bending actuation.
Fast carbonylation reaction from CO2 using plasma gas/liquid microreactors for radiolabeling applications
Laboratoire Procédés - Plasmas - Microsystèmes - Marion Gaudeau, Mengxue Zhang, Michaël Tatoulian, Camille Lescot and Stéphanie Ognier
Reaction Chemistry & Engineering - 5 1981-1991 - doi.org/10.1039/D0RE00289E. - 2020
Carbon-11 is undoubtedly an attractive PET radiolabeling synthon because carbon is present in all biological molecules. It is mainly found under 11CO2, but the latter being not very reactive, it is necessary to convert it into a secondary precursor. 11CO is an attractive precursor for labeling the carbonyl position through transition-metal mediated carbonylation because of its access to a wide range of functional groups (e.g., amides, ureas, ketones, esters, and carboxylic acids) present in most PET tracer molecules. However, the main limitations of 11CO labeling are the very short half-life of the radioisotope carbon-11 and its low concentration, and the low reactivity and poor solubility of 11CO in commonly used organic solvents. In this work, we show that a possible solution to these limitations is to use microfluidic reactor technology to perform carbonylation reactions, whilst a novel approach to generate CO from CO2 by plasma is described. The methodology consists of the decomposition of CO2 into CO by non-thermal DBD plasma at room temperature and atmospheric pressure, followed by the total incorporation of CO thus formed in the gas phase by carbonylation reaction, in less than 2 min of residence time. This “proof of principle” developed in carbon-12 would be further applied in carbon-11. Although considerable advances in 11CO chemistry have been reported in recent years, its application in PET tracer development is still an area of work in progress, because of the lack of commercially available synthesis instruments designed for 11C-carbonylations. To the best of our knowledge, such an innovative and efficient process, combining microfluidics and plasma, allowing the very fast organic synthesis of carbonyl molecules from CO2 with high yield, in mild conditions, has never been studied.
Plasma deposited high density amines on surface using (3- aminopropyl)triethoxysilane for assembling particles at sub-nano size
Laboratoire Procédés - Plasmas - Microsystèmes - Xi Rao , Ali Abou Hassan , Cédric Guyon , Stephanie Ognier , Michaël Tatoulian
Reaction Chemistry & Engineering - - DOI: 110.1016/j.matchemphys.2019.121974 - 2020
Although solid particles assembling on substrate surface is one of the key points for
developing membrane reactors, the technology of organizing nano/sub nanometer building
blocks into complex structures is still a challenge to scientists in years. In this work, amine
functional groups were deposited on the surface of different substrates via plasma enhanced
chemical vapor deposition (PECVD) technology and (3-aminopropyl)triethoxysilane
monomers were used as precursors. The influence of active gas, substrates, as well as
deposition time on the physico-chemical features of as-deposited film were investigated,
respectively. The highest density of amine of 5.5% on surface was obtained when Ar was
utilized as active gas and deposition time was 40 s. Furthermore, Y type zeolite particles at
sub-nano size were synthesized and subsequently used as a model material for testing the
immobilizing ability of plasma treated surface. The results clearly confirmed that a dense
mono or multi-layer of closely packed zeolite particles could be formed on the APTES as-
deposited surface after 24 hours’ immersion and the surface area of substrate could be
improved by the deposition of zeolite.
Amination of Cyclohexane by Dielectric Barrier Discharge Processing in a Continuous Flow Microreactor: Experimental and Simulation Studies
Laboratoire Procédés - Plasmas - Microsystèmes - Aurélien Lepoetre, Stéphanie Ognier, Mengxue Zhang, Julien Wengler, Safwan Al Ayoubi, Cyril Ollivier, Louis Fensterbank, Xavier Duten, Michael Tatoulian
Plasma Chemistry and Plasma Processing - - DOI: 10.1007/s11090-020-10140-9 - 2020
A miniaturized flow device has been developed to combine microfluidics technology and plasma process. In this microreactor, atmospheric pressure dielectric barrier discharges are generated in a gas in contact with a liquid phase. This study was conducted with plasma generated in ammonia in contact with a flow of liquid cyclohexane. Cyclohexylamine was synthesized with a good selectivity, and the process can be implemented to improve conversion and effectiveness. Numerical simulations confirmed that NH2 radicals are generated in the plasma and react with cyclohexyls radicals to achieve the reaction, giving a selectivity of 50% and a total molar conversion of 20% of cyclohexane. The effects of voltage and frequency on the selectivity and the experimental conversion rate were studied and discussed.
Using an Untargeted Metabolomics Approach to Identify Salivary Metabolites in Women with Breast Cancer
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Daniele Xavier Assad ,Ana Carolina Acevedo, Elisa Cançado Porto Mascarenhas, Ana Gabriela Costa Normando,Valérie Pichon,Helene Chardin,Eliete Neves Silva Guerra and Audrey Combes
Metabolites - 10(12) 1727-1751 - doi.org/10.3390/metabo10120506 - 2020
Metabolic alterations are a hallmark of the malignant transformation in cancer cells, which is characterized by multiple changes in metabolic pathways that are linked to macromolecule synthesis. This study aimed to explore whether salivary metabolites could help discriminate between breast cancer patients and healthy controls. Saliva samples from 23 breast cancer patients and 35 healthy controls were subjected to untargeted metabolomics using liquid chromatography-quadrupole time-of-flight mass spectrometry and a bioinformatics tool (XCMS Online), which revealed 534 compounds, characterized by their retention time in reverse-phase liquid chromatography and by the m/z ratio detected, that were shared by the two groups. Using the METLIN database, 31 compounds that were upregulated in the breast cancer group (p < 0.05) were identified, including seven oligopeptides and six glycerophospholipids (PG14:2, PA32:1, PS28:0, PS40:6, PI31:1, and PI38:7). In addition, pre-treatment and post-treatment saliva samples were analyzed for 10 patients who experienced at least a partial response to their treatment. In these patients, three peptides and PG14:2 were upregulated before but not after treatment. The area under the curve, sensitivity, and specificity for PG14:2 was 0.7329, 65.22%, and 77.14%, respectively. These results provide new information regarding the salivary metabolite profiles of breast cancer patients, which may be useful biomarkers. View Full-Text




Synthesis and characterization of molecularly imprinted polymers for the selective extraction of oxazepam from complex environmental and biological samples
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Daniele Xavier Assad Elisa Cançado Porto Mascarenhas Ana Gabriela Costa Normando Hélène Chardin Gustavo Barcelos Barra Riccardo Pratesi Yanna Karla De Medeiros Nóbrega Ana Carolina Acevedo Eliete Neves Silva Guerra
Molecular and Clinical Oncology - 155-161 - doi.org/10.3892/mco.2020.2062 - 2020
The early detection of breast cancer enables the use of less aggressive treatment and increases patient survival. The transmembrane glycoprotein mucin 1, which is also known as cancer antigen 15‑3 (CA15‑3), is aberrantly glycosylated and overexpressed in a variety of epithelial cancers, and serves a crucial role in the progression of the disease. CA15‑3 is currently used as a marker of breast cancer. In the present study, CA15‑3 concentrations in saliva and blood of patients with breast cancer were evaluated to test new assays to detect salivary CA15‑3 in addition to ELISA and its diagnostic value. To the best of our knowledge, there are no previous reports of the use of chemiluminescence assay (CLIA) and electrochemiluminescence assay (ECLIA) in saliva. Saliva and blood were collected on the same day from patients with breast cancer (n=26) and healthy controls (n=28). For each subject, the level of serum CA15‑3 was measured using ECLIA, and the level of salivary CA15‑3 was measured using ECLIA, CLIA and enzyme‑linked immunosorbent assay (ELISA). ELISA and CLIA were able to detect CA15‑3 in saliva; however, ECLIA could not detect salivary CA15‑3. There was no significant difference between the mean serum and salivary CA15‑3 levels in patients with breast cancer or healthy controls. The levels of CA15‑3 were highest for luminal breast cancer subtypes and stage IV cases. A moderate correlation was observed between salivary and serum CA15‑3 levels as measured by ELISA in breast cancer patients (r=0.56; P=0.0047). The results demonstrated that ECLIA was not a good method to detect salivary CA15‑3, although it is the gold standard for detecting serum CA15‑3. The presence of CA15‑3 in saliva was confirmed, and this will be useful in future research. Further investigations are necessary to confirm the ability to detect salivary CA15‑3 and its correlation with serum CA15‑3.
How high resolution mass spectrometry can help for the accurate quantification of difficult fragrance allergens
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Pierre‐Alain Remy, C. Pérès, J. Dugay, E. Corbi, Nathalie David, J. Vial
Flavour and Fragrance Journal - 36(1) - DOI:10.1002/ffj.3639 - 2020
Two high‐resolution mass spectrometers (HRMS) with different analyzer technology, Orbitrap and hybrid quadrupole time‐of‐flight (QTOF), were compared with a low‐resolution mass spectrometer, quadrupole, to analyse a set of 35 difficult allergens. These difficult allergens are commonly coeluted fragrance allergens with matrix compounds, using standard gas chromatography‐mass spectrometer conditions, from the extended list of the Scientific Committee on Consumer Safety (SCCS). Although the fundamental role of chromatographic separation has been demonstrated many times, the aim of this work is to demonstrate the benefits of high‐resolution. The added value of high‐resolution was illustrated in both a qualitative and a quantitative way. For qualitative aspect, the high resolution extracted ion signals of these two detectors were compared with the low‐resolution extracted ion signals. About 50% of the coeluted cases observed with the low‐resolution detector are easily resolved by the two high‐resolution detectors. For the quantitative aspect, an accuracy profile methodology and a performance metric were used to propose an overall evaluation. The Orbitrap mass spectrometer demonstrated a better overall performance, while the QTOF presented similar or even lower quantification performances than the quadrupole on the set of analysed fragrance
Impact of the Oil Matrix on Anionic and Nonionic Surfactant Separation Using Ultra-High-Performance Liquid Chromatography Hyphenated to High-Resolution Mass Spectrometry
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Alizée Dufour, Didier Thiébaut, Matthieu Loriau, Leticia Ligiero, and Jérôme Vial
American Chemical Society - 34(11) 13943–13953 - doi.org/10.1016/j.jchromb.2020.122518 - 2020
For the first time, to our knowledge, anionic and nonionic surfactants were analyzed in an oil matrix by ultra-high-performance liquid chromatography hyphenated to high-resolution mass spectrometry (UHPLC-HRMS). The feasibility of this analysis was studied using synthetic mixtures of surfactants prepared in water (quality controls), binary THF/toluene 50/50 v/v (surfactant + THF/toluene), and binary THF/toluene containing 1 and 10% crude oil (Crude1% and Crude10%). These compositions were chosen in order to be as close as possible to petroleum related samples to be investigated in the future. Analyses were carried out by UHPLC methods using both reverse phase and anion-exchange mechanisms with a mixed mode column. Despite the complexity of the oil matrix and the presence of organic solvents used for dilution, the retention times of the surfactants were not affected whatever the concentration of crude oil present in the sample. Nevertheless, a significant matrix effect caused a loss of signal when the concentration of oil reached 10% in mass. For the analysis of samples with this crude oil concentration range, it would be advisable to dilute the sample.




Performance evaluation of a MIP for the MISPE-LC determination of p-[18F]MPPF and a potential metabolite in human plasma
Laboratoire Spectrométrie de masse biologique et protéomique - F.Lecomte J.Aerts Plenevaux .Defraiteur. Chapuis-Hugonc. Rozetd. Chiape. Luxen. Pichon, Ph.Huberta C.Huberta
ELSEVIER - 180 113015 - https://doi.org/10.1016/j.jpba.2019.113015 - 2020
Within the family of serotonin (5-HT) receptors, the 5-HT1A subtype is particularly interesting as it may be involved in various physiological processes or psychological disorders. The p-[18F]MPPF, a highly selective 5-HT1A antagonist, is used for in vivo studies in human or animal by means of positron emission tomography (PET) [1].
In order to selectively extract p-[18F]MPPF and its main metabolites from plasma, molecularly imprinted polymer (MIP) was prepared against these compounds by using the p-MPPF as template. For the control of the selectivity, non-imprinted polymer (NIP) was also synthesized without template. The MIP sorbent, packed in disposable extraction cartridges (DECs), was then evaluated as molecularly imprinted solid-phase extraction (MISPE) prior to the LC determination. The conditions of extraction were evaluated in order to obtain the highest selective retention of the p-[18F]MPPF and its metabolites on this MIP. The MIP selectivity was exploited in the loading and washing steps by adjusting the pH of plasma samples at a suitable value and by selecting mixtures for the washing step to limit the contribution of non-specific interactions. Other important parameters involved in the conditioning and elution steps were also studied. Finally, a pre-validation was carried out with optimal extraction conditions to demonstrate the performance of this MISPE-LC method as a generic method in the context of evaluation of new MISPE for p-[18F]MPPF and its potential for metabolites extraction from human plasma.
Identification and semi-relative quantification of intact glycoforms by nano-LC–(Orbitrap)MS: application to the α-subunit of human chorionic gonadotropin and follicle-stimulating hormone
Laboratoire Spectrométrie de masse biologique et protéomique - Amira Al Matari, Audrey Combès, Julien Camperi, Thierry Fournier, Valérie Pichon & Nathalie Delaunay
ELSEVIER - 412 5729–5741 - , 10.1007/s00216-020-02794-3 - 2020
Human chorionic gonadotropin (hCG) and follicle-stimulating hormone (FSH) belong to the family of glycoprotein polypeptide hormones called gonadotropins. They are heterodimers sharing the α-subunit structure that has 2 N-glycosylation sites. A method based on nano-reversed-phase liquid chromatography coupled to high-resolution mass spectrometry with an Orbitrap analyzer was developed for the first time to characterize the glycosylation state of the α-subunit at the intact level. A recombinant hCG-based drug, Ovitrelle®, was analyzed. This method combined with an appropriate data treatment allowed the detection of not only the major isoforms but also the minority ones with a high mass accuracy. More than 30 hCGα glycoforms were detected without overlapping of the isotopic patterns. The figures of merit of the method were assessed. The relative standard deviations (RSDs) of the retention time ranged between 0.1 and 6.08% (n = 3), with an average of 0.4%. The RSDs of the peak area measured on the extracted ion chromatogram of each glycoform are below 38% (n = 3), with an average of 16%, thus allowing semi-relative quantification. The ability to accurately profile glycosylated variants of hCGα was next demonstrated by comparing qualitatively and semi-quantitatively 3 batches of Ovitrelle®. The method was also used to analyze 3 batches of a recombinant FSH-based drug, Puregon®, and 30 FSHα glycoforms were detected and semi-quantified. This demonstrates the high potential of this method for fast quality control or comparison of the glycosylation of glycoprotein-based pharmaceutical preparations.
Identification and semi-relative quantification of intact glycoforms by nano-LC–(Orbitrap)MS: application to the α-subunit of human chorionic gonadotropin and follicle-stimulating hormone
Laboratoire Spectrométrie de masse biologique et protéomique - Julien Camperi, Audrey Combès, Thierry Fournier, Valerie Pichon & Nathalie Delaunay
Research Paper - 412 4423–4432 - 10.1007/s00216-020-02684-8 - 2020
In the present work, the human chorionic gonadotropin (hCG) hormone was characterized for the first time by hydrophilic interaction liquid chromatography (HILIC) coupled to high-resolution (HR) quadrupole/time-of-flight (qTOF) mass spectrometry (MS) at the intact level. This heterodimeric protein, consisting of two subunits (hCGα and hCGβ), possesses 8 potential glycosylation sites leading to a high number of glycoforms and has a molecular weight of about 35 kDa. The HILIC conditions optimized in a first paper but using UV detection were applied here with MS for the analysis of two hCG-based drugs, a recombinant hCG and a hCG isolated from the urine of pregnant women. An amide column (150 × 2.1 mm, 2.6 μm, 150 Å), a mobile phase composed of acetonitrile and water both containing 0.1% of trifluoroacetic acid, and a temperature of 60 °C were used. The gradient was from 85 to 40% ACN in 30 min. The use of TFA that had been shown to be necessary for the separation of glycoforms caused, as expected, an ion suppression effect in MS that was partially overcome by increasing the amount of protein injected (2 μL at 1 mg mL−1) and reducing the detection m/z range (from 1500 to 300). These conditions allowed the detection of different glycoforms of hCGα. The performance of the HILIC-HRMS method was compared with that previously obtained in RPLC-HRMS in terms of the number of detected glycoforms, selectivity, and sensitivity. The complementarity and orthogonality of the HILIC and RP modes for the analysis of hCG at the intact level were demonstrated.
S-nitrosylation affects TRAP1 structure and ATPase activity and modulates cell response to apoptotic stimuli
Laboratoire Spectrométrie de masse biologique et protéomique - Fiorella Faienza, Matteo Lambrughi, Salvatore Rizza, Chiara Pecorari, Paola Giglio, Juan Salamanca Viloria, Maria Francesca Allega, Giovanni Chiappetta, Joëlle Vinh, Francesca Pacello, Andrea Battistoni, Andrea Rasola, Elena Papaleo, Giuseppe Filomeni
bioRxiv - - doi: 10.1016/j.bcp.2020.113869 - 2020
The mitochondrial chaperone TRAP1 has been involved in several mitochondrial functions, and modulation of its expression/activity has been suggested to play a role in the metabolic reprogramming distinctive of cancer cells. TRAP1 posttranslational modifications, i.e. phosphorylation, can modify its capability to bind to different client proteins and modulate its oncogenic activity. Recently, it has been also demonstrated that TRAP1 is S-nitrosylated at Cys501, a redox modification associated with its degradation via the proteasome. Here we report molecular dynamics simulations of TRAP1, together with analysis of long-range structural communication, providing a model according to which Cys501 S-nitrosylation induces conformational changes to distal sites in the structure of the protein. The modification is also predicted to alter open and closing motions for the chaperone function. By means of colorimetric assays and site directed mutagenesis aimed at generating C501S variant, we also experimentally confirmed that selective S-nitrosylation of Cys501 decreases ATPase activity of recombinant TRAP1. Coherently, C501S mutant was more active and conferred protection to cell death induced by staurosporine. Overall, our results provide the first in silico, in vitro and cellular evidence of the relevance of Cys501 S-nitrosylation in TRAP1 biology.
The zoonotic pathogen Leptospira interrogans mitigates environmental stress through cyclic-di-GMP-controlled biofilm production
Laboratoire Spectrométrie de masse biologique et protéomique - Thibeaux R, Soupé-Gilbert ME, Kainiu M, Girault D, Bierque E, Fernandes J, Bähre H, Douyère A, Eskenazi N, Vinh J, Picardeau M, Goarant C
NPJ Biofilms Microbiomes - 6(1) 24 - DOI [DOI] – 10.1038/s41522-020-0134-1 - 2020
The zoonotic bacterium Leptospira interrogans is the aetiological agent of leptospirosis, a re-emerging infectious disease that is a growing public health concern. Most human cases of leptospirosis result from environmental infection. Biofilm formation and its contribution to the persistence of virulent leptospires in the environment or in the host have scarcely been addressed. Here, we examined spatial and time-domain changes in biofilm production by L. interrogans. Our observations showed that biofilm formation in L. interrogans is a highly dynamic process and leads to a polarized architecture. We notably found that the biofilm matrix is composed of extracellular DNA, which enhances the biofilm’s cohesiveness. By studying L. interrogans mutants with defective diguanylate cyclase and phosphodiesterase genes, we show that biofilm production is regulated by intracellular levels of bis-(3′-5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) and underpins the bacterium’s ability to withstand a wide variety of simulated environmental stresses. Our present results show how the c-di-GMP pathway regulates biofilm formation by L. interrogans, provide insights into the environmental persistence of L. interrogans and, more generally, highlight leptospirosis as an environment-borne threat to human health.
On-Chip Sample Preparation Using a ChipFilter Coupled to NanoLC-MS/MS for Bottom-Up Proteomics
Laboratoire Spectrométrie de masse biologique et protéomique - Massamba M Ndiaye, Ha Phuong Ta, Giovanni Chiappetta, Joëlle Vinh
J Proteome Res - 19(7) 2654-2663 - DOI: 10.1021/acs.jproteome.9b00832 - 2020
Sample preparation is a crucial step in bottom-up proteomics. Analytical performances of bottom-up proteomics can be improved by the miniaturization of sample preparation. Many microfluidic devices have been designed in the field of proteomics, but many of them are not capable of handling complex samples and do not integrate the processing and digestion steps. We propose a ChipFilter Proteolysis (CFP) microfluidic device as a proteomics reactor for the miniaturization of protein sample processing and digestion steps, whose design is closely related to the experimental setup of filter-aided sample processing, even if no denaturing surfactant is required. The microchip has two reaction chambers of 0.6 μL volume separated by a protein filtration membrane in regenerated cellulose (10kD cutoff) that will concentrate or retain large polypeptides and will release small molecules. Cell lysis, protein concentration, and rapid chemical or enzymatic treatment can be performed in the ChipFilter. Complex proteomic samples like yeast protein extract or whole human cells proteome have been successfully analyzed with our microchip. Compared with the membrane-based commercial ultracentrifugation cartridge, our microfluidic device offered a better proteome coverage with 10 times less starting material and 8 times faster protocol duration.
Peptides Derived From Insulin Granule Proteins Are Targeted by CD8+ T Cells Across MHC Class I Restrictions in Humans and NOD Mice
Laboratoire Spectrométrie de masse biologique et protéomique - Marie Eliane Azoury, Mahmoud Tarayrah, Georgia Afonso, Aurore Pais, Maikel L. Colli, Claire Maillard, Cassandra Lavaud, Laure Alexandre-Heymann, Sergio Gonzalez-Duque, Yann Verdier4, Joelle Vinh, Sheena Pinto, Soren Buus, Danièle Dubois-L
Diabetes - 69(12) 2678-2690 - https://doi.org/10.2337/db20-0013 - 2020
The antigenic peptides processed by β-cells and presented through surface HLA class I molecules are poorly characterized. Each HLA variant (e.g., the most common being HLA-A2 and HLA-A3) carries some peptide-binding specificity. Hence, features that, despite these specificities, remain shared across variants may reveal factors favoring β-cell immunogenicity. Building on our previous description of the HLA-A2/A3 peptidome of β-cells, we analyzed the HLA-A3–restricted peptides targeted by circulating CD8+ T cells. Several peptides were recognized by CD8+ T cells within a narrow frequency (1–50/106), which was similar in donors with and without type 1 diabetes and harbored variable effector/memory fractions. These epitopes could be classified as conventional peptides or neoepitopes, generated either via peptide cis-splicing or mRNA splicing (e.g., secretogranin-5 [SCG5]–009). As reported for HLA-A2–restricted peptides, several epitopes originated from β-cell granule proteins (e.g., SCG3, SCG5, and urocortin-3). Similarly, H-2Kd–restricted CD8+ T cells recognizing the murine orthologs of SCG5, urocortin-3, and proconvertase-2 infiltrated the islets of NOD mice and transferred diabetes into NOD/scid recipients. The finding of granule proteins targeted in both humans and NOD mice supports their disease relevance and identifies the insulin granule as a rich source of epitopes, possibly reflecting its impaired processing in type 1 diabetes
Deciphering shell proteome within different Baltic populations of mytilid mussels illustrates important local variability and potential consequences in the context of changing marine conditions
Laboratoire Spectrométrie de masse biologique et protéomique - JaisonArivalagan, Benjamin Marie, Giovanni Chiappetta, Joëlle Vinh, Xavier Gallet, Matthieu Lebon Saloua M'Zoudi Philippe Dubois Sophie Berland Arul Marie
ELSEVIER - 745 140878 - doi.org/10.1016/j.scitotenv.2020.140878 - 2020
Molluscs defend themselves against predation and environmental stressors through the possession of mineralized shells. Mussels are widely used to predict the effects of abiotic factors such as salinity and pH on marine calcifiers in the context of changing ocean conditions. Shell matrix proteins are part of the molecular control regulating the biomineralization processes underpinning shell production. Under changing environmental conditions, differential expression of these proteins leads to the phenotypic plasticity of shells seen in many mollusc species. Low salinity decreases the availability of calcium and inorganic carbon in seawater and consequently energetic constraints often lead to thin, small and fragile shells in Mytilid mussels inhabiting Baltic Sea. To understand how the modulation of shell matrix proteins alters biomineralization, we compared the shell proteomes of mussels living under full marine conditions in the North Sea to those living in the low saline Baltic Sea. Modulation of proteins comprising the Mytilus biomineralization tool kit is observed. These data showed a relative increase in chitin related proteins, decrease in SD-rich, GA-rich shell matrix proteins indicating that altered protein scaffolding and mineral nucleation lead to impaired shell microstructures influencing shell resistance in Baltic Mytilid mussels. Interestingly, proteins with immunity domains in the shell matrix are also found to be modulated. Shell traits such as periostracum thickness, organic content and fracture resistance qualitatively correlates with the modulation of SMPs in Mytilid mussels providing key insights into control of biomineralization at molecular level in the context of changing marine conditions.

638 publications.