A5055496731- Protein Structure and Dynamics
- Advanced NMR Techniques and Applications
- Enzyme Structure and Function
- NMR spectroscopy and applications
- Molecular spectroscopy and chirality
- Electron Spin Resonance Studies
- Spectroscopy and Quantum Chemical Studies
- Advanced MRI Techniques and Applications
- RNA and protein synthesis mechanisms
- Lanthanide and Transition Metal Complexes
- RNA modifications and cancer
- Metabolomics and Mass Spectrometry Studies
- Heat shock proteins research
- RNA Research and Splicing
- Mass Spectrometry Techniques and Applications
- DNA Repair Mechanisms
- Glycosylation and Glycoproteins Research
- Biochemical and Structural Characterization
- Protein Kinase Regulation and GTPase Signaling
- Melanoma and MAPK Pathways
- Microbial Natural Products and Biosynthesis
- Influenza Virus Research Studies
- Chemical Synthesis and Analysis
- Cancer Mechanisms and Therapy
- Computational Drug Discovery Methods
Centre National de la Recherche Scientifique
2015-2025
Sorbonne Université
2016-2025
Université Paris Sciences et Lettres
2016-2025
École Normale Supérieure
2023-2025
École Normale Supérieure - PSL
2016-2025
Laboratoire des Biomolécules
2016-2024
Institut Superieur de l'Aeronautique et de l'Espace (ISAE-SUPAERO)
2016
Institut de Biologie Structurale
2005-2015
Université Grenoble Alpes
2005-2015
CEA Grenoble
2006-2015
Ever since its initial development, solution NMR spectroscopy has been used as a tool to study conformational exchange. Although many systems are amenable relaxation dispersion approaches, cases involving highly skewed populations in slow chemical exchange have, general, remained recalcitrant study. Here an experiment detect and characterize "invisible" excited protein states with visible ground-state conformation (excited-state lifetimes ranging from ∼5 50 ms) is presented. This method,...
Despite their importance for biological activity, slower molecular motions beyond the nanosecond range remain poorly understood. We have assembled an unprecedented set of experimental NMR data, comprising up to 27 residual dipolar couplings per amino acid, define nature and amplitude backbone motion in protein G using Gaussian axial fluctuation model three dimensions. Slower occur loops, beta-sheet, are absent other regions molecule, including alpha-helix. In beta-sheet alternating pattern...
Biological function relies on the complex spectrum of conformational dynamics occurring in biomolecules. We have combined Accelerated Molecular Dynamics (AMD) with experimental results derived from NMR to probe multiple time-scale motions third IgG-binding domain Protein G (GB3). AMD is shown accurately reproduce amplitude and distribution slow motional modes characterized using residual dipolar couplings, reporting up millisecond timescale. In agreement experiment, larger slower are...
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease involving cytotoxic conformations of Cu, Zn superoxide dismutase (SOD1). A major challenge in understanding ALS pathology has been the identification and atomic-level characterization these conformers. Here, we use combination NMR methods to detect four distinct sparsely populated transiently formed thermally accessible conformers equilibrium with native state immature SOD1 (apoSOD12SH). Structural models two...
The 70-kDa heat shock protein (Hsp70) family of chaperones bind cognate substrates to perform a variety different processes that are integral cellular homeostasis. Although detailed structural information is available on the chaperone, features folding competent in bound form have not been well characterized. Here we use paramagnetic relaxation enhancement (PRE) NMR spectroscopy probe existence long-range interactions one such substrate, human telomere repeat binding factor (hTRF1), which...
Abstract Aromatic residues cluster in the core of folded proteins, where they stabilize structure through multiple interactions. Nuclear magnetic resonance (NMR) studies 1970s showed that aromatic side chains can undergo ring flips—that is, 180° rotations—despite their role maintaining protein fold 1–3 . It was suggested large-scale ‘breathing’ motions surrounding environment would be necessary to accommodate these flipping events 1 However, structural details have remained unclear. Here we...
Abstract. Combining high-field high-resolution NMR with an evolution of spin systems at low magnetic field offers many opportunities for the investigation molecular motions, hyperpolarization and exploration field-dependent dynamics. Fast reproducible transfer between high fields is required to minimize polarization losses due longitudinal relaxation. Here, we introduce a new design sample shuttle that achieves remarkably speeds, vmax ~ 27 m.s-1. This hybrid pneumatic/mechanical apparatus...
An atomic resolution description of protein flexibility is essential for understanding the role that structural dynamics play in biological processes. Despite unique dependence nuclear magnetic resonance (NMR) to motional averaging on different time scales, NMR-based structure determination often ignores presence dynamics, representing rapidly exchanging conformational equilibria terms a single static structure. In this study, we use rich dynamic information encoded experimental NMR...
The 70 kDa heat shock protein (Hsp70) chaperone system is ubiquitous, highly conserved, and involved in a myriad of diverse cellular processes. Its function relies on nucleotide-dependent interactions with client proteins, yet the structural features folding-competent substrates their Hsp70-bound state remain poorly understood. Here we use NMR spectroscopy to study human telomere repeat binding factor 1 (hTRF1) complex Escherichia coli Hsp70 (DnaK). In complex, hTRF1 globally unfolded up 40%...
Molecular recognition is integral to biological function and frequently involves preferred binding of a molecule one several exchanging ligand conformations in solution. In such process the bound structure can be selected from ensemble interconverting ligands priori (conformational selection, CS) or may form once (induced fit, IF). Here we focus on ubiquitous conserved Hsp70 chaperone which oversees integrity cellular proteome through its ATP-dependent interaction with client proteins. We...
Chemical exchange saturation transfer (CEST) NMR spectroscopy has emerged as a powerful technique for studies of transiently formed, sparsely populated (excited) conformational states protein molecules in slow with dominant structure. The most popular form the experiment, and version originally developed, uses weak (1)H radio frequency field to perturb longitudinal magnetization one state effect transferred second conformation via chemical exchange. A significant limitation method...
Intrinsically disordered proteins (IDPs) display a large number of interaction modes including folding-upon-binding, binding without major structural transitions, or through highly dynamic, so-called fuzzy, complexes. The vast majority experimental information about IDP have been inferred from crystal structures in complex with short peptides IDPs. However, provide mainly static view the complexes and do not give conformational dynamics experienced by bound state. Knowledge is fundamental...
Significance In the mitogen-activated protein kinase (MAPK) pathways, N-terminal intrinsically disordered regulatory domains of MAPK kinases (MKK) control signaling specificity by binding to their cognate MAPKs via docking sites carrying homologous recognition sequences. MKK7 activates c-Jun (JNK) pathway and is only MKK containing three motifs within its domain. Here we obtain a comprehensive picture structure, dynamics, affinity, stoichiometry, kinetics MKK7–JNK complex. Importantly, show...
A robust procedure for the determination of protein-backbone motions on time scales pico- to milliseconds directly from residual dipolar couplings has been developed that requires no additional scaling relative external references. The results ubiquitin (blue in graph: experimental N-HN order parameters) correspond closely amplitude, nature, and distribution motion found a 400 ns molecular-dynamics trajectory (red).
Influenza A RNA polymerase complex is formed from three components, PA, PB1, and PB2. PB2 independently imported into the nucleus prior to reconstitution. All crystallographic structures of C-terminus (residues 536-759) reveal two globular domains, 627 NLS, that form a tightly packed heterodimer. The molecular basis affinity 627-NLS for importins remained unclear these structures, apparently requiring large-scale conformational changes importin binding. Using combination solution-state NMR,...
Determination of protein structure classically results in a single average configuration that takes no account conformational fluctuation. Dynamics are, however, inherently linked to and crucial our understanding biological function. In this study we have used analytical descriptions dynamic averaging residual dipolar couplings (RDCs) simultaneously determine the backbone dynamics GB3. RDCs alone are an ultrahigh-resolution compares very closely with refined X-ray (rmsd 0.34 Å overall...
A pulse scheme is presented for quantifying millisecond time scale chemical exchange processes in proteins by measuring 1H CPMG relaxation dispersion profiles of 13CHD2 methyl groups. The use isotopomers experiments eliminates problems with interconversion between differentially relaxing proton transitions that complicate the extraction accurate parameters when 13CH3 probes are used. Good agreement demonstrated extracted shift differences from fits and corresponding measured independently on...
Carr-Purcell-Meiboom-Gill relaxation dispersion NMR spectroscopy has emerged as a valuable tool to characterize conformational exchange between major and minor states in large variety of biomolecules. The window that is amenable for study, corresponding rates on the order 2000 s(-1) or less, limiting, however. Here we show combined analysis both amide (15)N (1)H(N) CPMG profiles state induced chemical shift changes leads significant increases time scale which accurate parameters differences...
Significance Site-specific rates of exchange amide hydrogens with solvent protons are important parameters for characterizing protein structure and dynamics, providing insight into a range biomolecular processes. In typical hydrogen experiments, measured reflect from all accessible states protein. Here we describe an NMR approach measuring in sparsely populated, transiently formed conformational that interconvert highly populated long-lived ground state. An application to folding reaction...
Chemical exchange saturation transfer (CEST) has recently evolved into a powerful approach for studying sparsely populated, "invisible" protein states in slow with major, visible conformer. Central to the technique is use of weak, highly selective radio-frequency field that applied at different frequency offsets successive experiments, "searching" minor state resonances. The recording CEST profiles enough points ensure coverage entire spectrum sufficient resolution can be time-consuming,...