Biophysics of Macromolecules and their Interactions - CNRS URA 2185  

  HEADDr. Patrick ENGLAND /
  MEMBERSBruno BARON / Sylviane HOOS / Bertrand RAYNAL / Jocelyne FRAYSSE

  Annual Report

Molecular-scale biophysical approaches are a privileged link between the atomic-level structural descriptions and the spatiotemporal in situ studies, providing invaluable information about the energetics and dynamics of biological macromolecules and assemblies in a relatively short time-scale and in close-to-physiological conditions.

The Centre of Biophysics of Macromolecules and their Interactions (french acronym: PFBMI) aims to group state-of-the-art equipment, adapted in priority to the needs of the research teams of the Pasteur Institute and its International Network, together with the corresponding technical and methodological know-how, thereby providing a scientific environment conducive to world-class research in the field of macromolecular science.

The following technologies are presently available(december 2008):

1) Analytical ultracentrifugation:ProteomeLab XL I & Optima XL-A ultracentrifuges (Beckman-Coulter).

2) Circular dichroism: Aviv 215 spectropolarimeter (Aviv Biomedical), with fluorescence and titration accessories ; CD6 spectropolarimeter (Jobin-Yvon) with a “stopped-flow” accessory (Bio-Logic).

3) Fluorescence spectroscopy:Quantamaster C60 fluorimeter (Photon Technology International), with a polarisation module ; KinetAsyst SF61-DX2 “stopped flow” instrument (Hi-Tech Scientific), with a polarisation module.

4) Infrared spectroscopy:MB104 spectrometer (ABB Bomem), with a DuraSamplIR II attenuated total reflectance accessory (SensIR).

5) Light scattering: DynaPro MS800 dynamic light scattering instrument (Wyatt) ; TDA 302 light scattering-intrinsic viscosity-refractive index triple detector array (Viscotek), coupled to a GPCmax size exclusion chromatography system.

6) Microcalorimetry: VP-ITC & MCS-ITC isothermal titration calorimeters ; VP-DSC differential scanning calorimeter, with a Pressure Perturbation Calorimetry accessory (PPC) (MicroCal).

7) Surface plasmon resonance:Biacore 2000 and Biacore X100 biosensor (Biacore), and ProteOn XPR36 protein interaction array system (Bio-Rad)

People wishing to use the instruments of the PFBMI, or seeking expert advice related to these technologies, must send an e-mail to a centralized address, Instructions and down-loadable forms are available from our Web site, as a support for these applications, which can be made at any time.

Experiments can be carried out according to 3 different schemes: 1) instrument allocation after user training; 2) service provision; 3) scientific collaboration.

In the past 4 years, the 15 instruments and the know-how of the PFBMI have attracted more than 120 projects altogether, in association with 30 Research Units of the Institut Pasteur (from all 10 Scientific Departments) and 25 laboratories from other institutions (french or foreign), leading to 42 peer-reviewed publications.

Below is a very small sample of subjects covered by scientific collaborations and having lead to publications since 2005:

* Characterization of the coupling between the trimerization of the rabies virus glycoprotein and its interaction with nerve growth factor receptor p75NTR(J. Gen. Virol. (2005) 86, 2543-2552)

* Determination of the effect of natural pathogenic mutations of the NF-κB modulator protein NEMO on its stability and oligomerization properties(J. Biol. Chem (2006) 281, 6334-6348)

* Study of the conformational changes induced by calcium in the RTX subdomain of Bordetella pertussis adenylate cyclase toxin J. Biol. Chem (2006) 281, 16914-16926; J. Biol. Chem (2009) 284, 1781-1789)

* Physico-chemical characterization of Sta1, a novel archaeal transcriptional activator(N.A.R. (2006) 34, 4837-4845)

* Study of the mechanism of action of transcriptional regulators TcdC from Clostridium difficile (Mol. Microbiol. (2007) 64, 1274–1288) and Crl from Escherichia coli (J. Biol. Chem. (2008) 283, 33455-33464)

* Structural and thermodynamical characterization of the properties of prolactin receptor antagonists (J. Biol. Chem (2007) 282, 33118-33131)

* Demonstration of the IgE binding capacity of murine receptor FcγRIV (J. Clin. Invest. (2008) 118, 3738-3750)

* Characterization of the mechanism of action of CymR, central regulator of the cysteine metabolism in Bacillus subtilis (J. Biol. Chem (2008) 283, 33561-33560)

Keywords: spectroscopy, hydrodynamics, thermodynamics, kinetics, proteins, polysaccharides, nucleic acids, lipids



England P, Westblade LF, Karimova G, Robbe-Saule V, Norel F & Kolb A (2008) "Binding of the unorthodox transcription activator, Crl, to the components of the transcription machinery". Journal of Biological Chemistry283, 33455-33464

Tanous C, Soutourina O, Raynal B, Hullo MF, Mervelet P, Gilles AM, Noirot P, Danchin A, England P& Martin-Verstraete I. (2008) . "The CymR regulator in complex with the enzyme CysK controls cysteine metabolism in Bacillus subtilis". Journal of Biological Chemistry283, 33551-33560

Jomain JB; Tallet E; Broutin I; Hoos S; Van Agthoven J; Kelly PA; Ducruix A; Kragelund BB; England P& Goffin V. (2007) "Structural and thermodynamical bases for the design of pure prolactin receptor antagonists: X-ray structure of Del1-9-G129R-hPRL" . Journal of Biological Chemistry282, 33118-33131

Matamouros S; England P& Dupuy B (2007) . "Clostridium difficile toxin expression is inhibited by the novel regulator TcdC"; Molecular Microbiology64, 1274–1288

Villarino A; Duran R; Wehenkel A; Fernandez P; England P; Brodin P; Cole ST; Zimny-Arnat U; Jungblut PR; Cerveñansky C & Alzari PM (2005) . "Proteomic identification of M. tuberculosis protein kinase substrates: PknB recruits GarA, a FHA domain-containing protein, through activation loop-mediated interactions". Journal of Molecular Biology 350, 953–963

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Activity Reports 2009 - Institut Pasteur
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