A5017194362- Mass Spectrometry Techniques and Applications
- Advanced Proteomics Techniques and Applications
- Mitochondrial Function and Pathology
- Metabolism and Genetic Disorders
- Glycosylation and Glycoproteins Research
- ATP Synthase and ATPases Research
- Monoclonal and Polyclonal Antibodies Research
- DNA Repair Mechanisms
- Analytical Chemistry and Chromatography
- Multiple Myeloma Research and Treatments
- Vibrio bacteria research studies
- Bacterial Genetics and Biotechnology
- Biotin and Related Studies
- Photosynthetic Processes and Mechanisms
- Genomics and Chromatin Dynamics
- Metabolomics and Mass Spectrometry Studies
- Amyloidosis: Diagnosis, Treatment, Outcomes
- Enzyme Structure and Function
- Photoreceptor and optogenetics research
- RNA and protein synthesis mechanisms
- Biochemical and Structural Characterization
- Celiac Disease Research and Management
- DNA and Nucleic Acid Chemistry
- Alzheimer's disease research and treatments
- Plant and Biological Electrophysiology Studies
Université Paris Cité
2022-2024
Institut Pasteur
2018-2024
Centre National de la Recherche Scientifique
2008-2024
University of Calgary
2011-2023
Sanofi (France)
2021
Heinrich Heine University Düsseldorf
2021
Technical University of Denmark
2021
Institut Textile et Chimique
2020
Commissariat à l'Énergie Atomique et aux Énergies Alternatives
2009-2012
Inserm
2011-2012
The XRCC4-like factor (XLF)-XRCC4 complex is essential for nonhomologous end joining, the major repair pathway DNA double strand breaks in human cells. Yet, how XLF binds XRCC4 and impacts joining functions has been enigmatic. Here, we report XLF-XRCC4 crystal structure combination with biophysical mutational analyses to define interactions. Crystal solution structures plus mutations characterize alternating head domain interfaces forming parallel super-helical filaments. Leu-115...
Hydrogen/deuterium exchange coupled to mass spectrometry (HXMS) utilizes enzymatic digestion of proteins localize the information about altered patterns in protein structure. The ability protease produce small peptides and overlapping fragments provide sufficient coverage sequence is essential for localizing regions interest. Recently, it was shown that there an interesting group proteolytic enzymes from carnivorous pitcher plants genus Nepenthes. In this report, we describe successful...
Abstract Celiac disease is triggered by partially digested gluten proteins. Enzyme therapies that complete protein digestion in vivo could support a gluten-free diet, but the barrier to completeness high. Current options require enzyme amounts on same order as meal itself. In this study, we evaluated proteolytic components of carnivorous pitcher plant ( Nepenthes spp.) for use context. Remarkably low doses enhance gliadin solubilization rates and degrade slurries within pH temporal...
Abstract Bacteria have evolved toxins to outcompete other bacteria or hijack host cell pathways. One broad family of bacterial polymorphic gathers multidomain proteins with a modular organization, comprising C-terminal toxin domain fused N-terminal that adapts the delivery apparatus. Polymorphic include bacteriocins, contact-dependent growth inhibition systems, and specialized Hcp, VgrG, PAAR Rhs Type VI secretion (T6SS) components. We recently described characterized Tre23, T6SS-associated...
Detergents are frequently used for protein isolation and solubilization. Their presence is crucial in membrane protocols or lipid raft proteomics. However, they usually poorly compatible with mass spectrometry. Several different sample preparation routinely used, but either laborious suffer from losses. Here, we describe our alternative method nonionic detergent removal. It based on selective extraction after capture of the a reversed phase cartridge. The performed by chlorinated solvents...
The pitcher secretions of the Nepenthes genus carnivorous plants contain a proteolytic activity that is very useful for hydrogen/deuterium exchange mass spectrometry (HX-MS). Our efforts to reconstitute fluid using recombinant nepenthesin I (one two known aspartic proteases in fluid) revealed partial cleavage profile and reduced enzymatic stability certain HX-MS applications. We produced characterized II determine if it complemented Nepenthesin shares many properties with I, such as fast...
Studies of protein dynamics, structure and interactions using hydrogen/deuterium exchange mass spectrometry (HDX-MS) have sharply increased over the past 5–10 years. The predominant technology requires fast digestion at pH 2–3 to retain deuterium label. Pepsin is used almost exclusively, but it provides relatively low efficiency under constraints experiment, a selectivity profile that renders poor coverage intrinsically disordered regions. In this study we present nepenthesin-containing...
Hydrogen/deuterium exchange (HDX) methods generate useful information on protein structure and dynamics, ideally at the individual residue level. Most MS-based HDX involve a rapid proteolytic digestion followed by LC/MS analysis, with kinetics monitored peptide Localizing specific sites of is usually restricted to resolution size host because gas-phase processes can scramble deuterium throughout peptide. Subtractive may improve resolution, where levels overlapping nested peptides are used in...
Non-homologous end joining (NHEJ) repairs DNA double strand breaks in non-cycling eukaryotic cells. NHEJ relies on polynucleotide kinase/phosphatase (PNKP), which generates 5΄-phosphate/3΄-hydroxyl termini that are critical for ligation by the ligase, LigIV. PNKP and LigIV require scaffolding protein, XRCC4. The FHA domain binds to CK2-phosphorylated XRCC4 C-terminal tail, while uses its tandem BRCT repeats bind coiled-coil. Yet, assembled PNKP-XRCC4-LigIV complex remains uncharacterized....
Trypsin dominates bottom-up proteomics, but there are reasons to consider alternative enzymes. Improving sequence coverage, exposing proteomic "dark matter," and clustering post-translational modifications in different ways with higher-order drive the pursuit of reagents complementary trypsin. Additionally, enzymes that easy use generate larger peptides capitalize upon newer fragmentation technologies should have a place proteomics. We expressed characterized recombinant neprosin, novel...
The current technique used for microbial identification in hospitals is matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). However, it suffers from important limitations, particular, closely related species or when the database lacks appropriate reference. In this work, we set up a liquid chromatography (LC)–MS/MS top-down proteomics platform, which aims at discriminating pathogenic bacteria through of specific proteoforms. Using Escherichia coli as...
Chemical cross-linking (XL) coupled to mass spectrometry (MS) has become a powerful approach probe the structure of protein assemblies. Although most applications concerned purified complexes, latest developments focus on large-scale in vivo studies. Pushing this direction, we developed an advanced platform study cellular interactome living bacterial cells. It is based labeling and involves one-step enrichment by click chemistry solid support. Our shows impressive efficiency Neisseria...
In multiple myeloma diseases, monoclonal immunoglobulin light chains (LCs) are abundantly produced, with, as a consequence in some cases, the formation of deposits affecting various organs, such kidney, while other cases remaining soluble up to concentrations several g·L–1 plasma. The exact factors crucial for solubility LCs poorly understood, but it can be hypothesized that their amino acid sequence plays an important role. Determining precise sequences patient-derived is therefore highly...
Abstract Type IV pili (T4P) are prevalent, polymeric surface structures in pathogenic bacteria, making them ideal targets for effective vaccines. However, bacteria have evolved efficient strategies to evade type pili-directed antibody responses. Neisseria meningitidis prototypical pili-expressing Gram-negative responsible life threatening sepsis and meningitis. This species has several genetic modify the of its pili, changing pilin subunit amino acid sequence, nature glycosylation...
Abstract Hydrogen/deuterium (H/D) exchange coupled to mass spectrometry is nowadays routinely used probe protein interactions or conformational changes. The method has many advantages, e.g. very low sample consumption, but offers limited spatial resolution. One way higher resolution leads through the use of different proteases their combinations. In present work we describe recombinant production, purification and aspartic protease zymogen from Rhizopus chimensis , type XVIII (EC 3.4.23.6),...
The mitochondrial adenine nucleotide carrier (Ancp) catalyzes the transport of ADP and ATP across inner membrane, thus playing an essential role in cellular energy metabolism. During mechanism switches between two different conformations that can be blocked by toxins: carboxyatractyloside (CATR) bongkrekic acid. Therefore, our understanding improved analyzing structural differences individual inhibited states. We have solved three-dimensional structure bovine isoform 1 (bAnc1p) a complex...
Hydrogen-deuterium exchange mass spectrometry is an important method for protein structure-function analysis. The bottom-up approach uses digestion to localize deuteration higher resolution, and the essential measurement involves centroid determinations on a very large set of peptides. In course evaluating systems various projects, we established two (HDX-MS) platforms that consisted FT-MS high-resolution QTOF spectrometer, each with matched front-end fluidic systems. Digests proteins...
The mitochondrial ADP/ATP carrier catalyzes the transport of ADP and ATP across inner membrane by switching between two different conformations. They can be blocked inhibitors: carboxyatractyloside (CATR) bongkrekic acid (BA). Our understanding process is largely based on analysis structural differences individual inhibited states. X-ray crystallographic three-dimensional structure bovine isoform 1 (bAnc1p) complexed with CATR was determined, but BA–carrier complex remains unknown. We...