CNRS Biomolecules Laboratory

Paris, France

CNRS Biomolecules Laboratory

Paris, France
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Calligari P.,CNRS Biomolecules Laboratory | Abergel D.,CNRS Biomolecules Laboratory
Journal of Physical Chemistry B | Year: 2012

In this article, we investigate the multiple-scale structure of methyl side chain dynamics in proteins. We show that the orientational correlation functions of CH3 methyl groups are well described by a fractional Brownian dynamics model. Typical angular correlation functions involved in NMR relaxation were computed from MD simulations performed on two different proteins. These correlation functions were shown to be very well fitted by a fractional Ornstein-Uhlenbeck process in the presence of effective local potentials at the C-H and C-C methyl bonds. In addition, our analysis highlights the presence of the asymptotic power law decay of the waiting time probability density of the stochastic process involved, thereby illustrating the connection between approaches based on fractional diffusion equations and the continuous time random walk. © 2012 American Chemical Society.

Caillon L.,CNRS Biomolecules Laboratory | Lequin O.,CNRS Biomolecules Laboratory | Khemtemourian L.,CNRS Biomolecules Laboratory
Biochimica et Biophysica Acta - Biomembranes | Year: 2013

Human islet amyloid polypeptide (IAPP) forms amyloid fibrils in the pancreatic islets of patients suffering from type 2 diabetes mellitus (T2DM). The formation of IAPP fibrils has been shown to cause membrane damage which most likely is responsible for the death of pancreatic islet β-cells during the pathogenesis of T2DM. Several studies have demonstrated a clear interaction between IAPP and lipid membranes. However the effect of different lipid compositions and of various membrane mimetics (including micelles, bicelles, SUV and LUV) on fibril formation kinetics and fibril morphology has not yet systematically been analysed. Here we report that the interaction of IAPP with various membrane models promoted different processes of fibril formation. Our data reveal that in SDS and DPC micelles, IAPP adopts a stable α-helical structure for several days, suggesting that the micelle models may stabilize monomeric or small oligomeric species of IAPP. In contrast, zwitterionic DMPC/DHPC bicelles and DOPC SUV accelerate the fibril formation compared to zwitterionic DOPC LUV, indicating that the size of the membrane model and its curvature influence the fibrillation process. Negatively charged membranes decrease the lag-time of the fibril formation kinetics while phosphatidylethanolamine and cholesterol have an opposite effect, probably due to the modulation of the physical properties of the membrane and/or due to direct interactions with IAPP within the membrane core. Finally, our results show that the modulation of lipid composition influences not only the growth of fibrils at the membrane surface but also the interactions of β-sheet oligomers with membranes. © 2013 Elsevier B.V.

Stafford K.A.,Columbia University | Ferrage F.,CNRS Biomolecules Laboratory | Cho J.-H.,Texas A&M University | Palmer A.G.,Columbia University
Journal of the American Chemical Society | Year: 2013

Many proteins use Asx and Glx (x = n, p, or u) side chains as key functional groups in enzymatic catalysis and molecular recognition. In this study, NMR spin relaxation experiments and molecular dynamics simulations are used to measure the dynamics of the side chain amide and carboxyl groups, 13Cγ/δ, in Escherichia coli ribonuclease HI (RNase H). Model-free analysis shows that the catalytic residues in RNase H are preorganized on ps-ns time scales via a network of electrostatic interactions. However, chemical exchange line broadening shows that these residues display significant conformational dynamics on μs-ms time scales upon binding of Mg2+ ions. Two groups of catalytic residues exhibit differential line broadening, implicating distinct reorganizational processes upon binding of metal ions. These results support the "mobile metal ion" hypothesis, which was inferred from structural studies of RNase H. © 2013 American Chemical Society.

Tekely P.,CNRS Biomolecules Laboratory
Solid State Nuclear Magnetic Resonance | Year: 2015

Combined use of cross-polarization and magic-angle spinning in the middle of the seventies has opened a new era of high-resolution solid-state NMR spectroscopy. Cross-polarization procedure is commonly used to obtain a shorter measuring time and to investigate or exploit one nucleous by means of the other nucleous involved in the polarization transfer. An extended family of cross-polarization experiments including constant time cross-polarization approach, cross-polarization inversion and indirect observation of proton spin system is reviewed and illustrated with applications to a large range of solids. © 2015 Elsevier Inc.

Calligari P.,International School for Advanced Studies | Abergel D.,CNRS Biomolecules Laboratory
Journal of Physical Chemistry B | Year: 2014

Fluctuations of NMR resonance frequency shifts and their relation with protein exchanging conformations are usually analyzed in terms of simple two-site jump processes. However, this description is unable to account for the presence of multiple time scale dynamics. In this work, we present an alternative model for the interpretation of the stochastic processes underlying these fluctuations of resonance frequencies. Time correlation functions of 15N amide chemical shifts computed from molecular dynamics simulations (MD) were analyzed in terms of a transiently fractional diffusion process. The analysis of MD trajectories spanning dramatically different time scales (∼200 ns and 1 ms [ Shaw, D. E.; Science 2010, 330, 341-346 ]) allowed us to show that our model could capture the multiple scale structure of chemical shift fluctuations. Moreover, the predicted exchange contribution Rex to the NMR transverse relaxation rate is in qualitative agreement with experimental results. These observations suggest that the proposed fractional diffusion model may provide significative improvement to the analysis of NMR dispersion experiments. © 2014 American Chemical Society.

Ferrage F.,CNRS Biomolecules Laboratory
Methods in Molecular Biology | Year: 2012

Nitrogen-15 relaxation is the most ubiquitous source of information about protein (backbone) dynamics used by NMR spectroscopists. It provides the general characteristics of hydrodynamics as well as internal motions on subnanosecond, micro- and millisecond timescales of a biomolecule. Here, we present a full protocol to perform and analyze a series of experiments to measure the 15N longitudinal relaxation rate, the 15N transverse relaxation rate under an echo train or a single echo, the 15N- 1H dipolar cross-relaxation rate, as well as the longitudinal and transverse cross-relaxation rates due to the cross-correlation of the nitrogen-15 chemical shift anisotropy and the dipolar coupling with the adjacent proton. These rates can be employed to carry out model-free analyses and can be used to quantify accurately the contribution of chemical exchange to transverse relaxation. © 2012 Springer Science+Business Media, LLC.

Kurzbach D.,CNRS Biomolecules Laboratory
Protein Science | Year: 2016

A methodological framework is presented for the graph theoretical interpretation of NMR data of protein interactions. The proposed analysis generalizes the idea of network representations of protein structures by expanding it to protein interactions. This approach is based on regularization of residue-resolved NMR relaxation times and chemical shift data and subsequent construction of an adjacency matrix that represents the underlying protein interaction as a graph or network. The network nodes represent protein residues. Two nodes are connected if two residues are functionally correlated during the protein interaction event. The analysis of the resulting network enables the quantification of the importance of each amino acid of a protein for its interactions. Furthermore, the determination of the pattern of correlations between residues yields insights into the functional architecture of an interaction. This is of special interest for intrinsically disordered proteins, since the structural (three-dimensional) architecture of these proteins and their complexes is difficult to determine. The power of the proposed methodology is demonstrated at the example of the interaction between the intrinsically disordered protein osteopontin and its natural ligand heparin. © 2016 The Protein Society.

Bechara C.,CNRS Biomolecules Laboratory | Sagan S.,CNRS Biomolecules Laboratory
FEBS Letters | Year: 2013

Twenty years ago, the discovery of peptides able to cross cellular membranes launched a novel field in molecular delivery based on these non-invasive vectors, most commonly called cell-penetrating peptides (CPPs) or protein transduction domains (PTDs). These peptides were shown to efficiently transport various biologically active molecules inside living cells, and thus are considered promising devices for medical and biotechnological developments. Moreover, CPPs emerged as potential tools to study the prime mechanisms of cellular entry across the plasma membrane. This review is dedicated to CPP fundamentals, with an emphasis on the molecular requirements and mechanism of their entry into eukaryotic cells. © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Burlina F.,CNRS Biomolecules Laboratory | Papageorgiou G.,National Insitiute for Medical Research | Morris C.,National Insitiute for Medical Research | White P.D.,Merck And Co. | Offer J.,National Insitiute for Medical Research
Chemical Science | Year: 2014

The progress of total chemical protein synthesis has been hampered by difficulties in preparing peptide thioesters by standard Fmoc peptide synthesis. The amino acid, α-methylcysteine, sited at the C-terminus of a peptide can substitute for a thioester in peptide ligation reactions. C-terminal α-methylcysteine is fully compatible with Fmoc peptide synthesis and its use in ligation is very simple and robust. Its potential is demonstrated with the synthesis of model proteins. © 2014 The Royal Society of Chemistry.

Walrant A.,CNRS Biomolecules Laboratory | Bechara C.,CNRS Biomolecules Laboratory | Alves I.D.,French National Center for Scientific Research | Sagan S.,CNRS Biomolecules Laboratory
Nanomedicine | Year: 2012

Cell-penetrating peptides are short basic peptide sequences that might display amphipathic properties. These positively charged peptides internalize into all cell types, albeit with different efficiency. Cell-penetrating peptides use all routes of pinocytosis to internalize, in addition to direct membrane translocation that requires interaction with lipid membrane domains. These differences in internalization efficiency according to the peptide sequence and cell type suggest that the cell-penetrating peptides interact with different molecular partners at the cell surface. This review will first report on data that describe the molecular interaction of the most popular cell-penetrating peptides (penetratin, Tat and oligoarginine) with carbohydrates and lipids. The second part of the review will be dedicated to cell studies that have reported how cell surface composition influences cell internalization. Discussion will focus on the gap between in vitro and in cellulo studies, and more specifically to which extent the interaction with molecules found in membranes reflect the internalization efficiency of the peptides. © 2012 Future Medicine Ltd.

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