Biophysics of Macromolecules and their Interactions - CNRS URA 2185  

  HEADDr. Patrick ENGLAND /
  MEMBERSEngineers: Dr. Bruno BARON / Sylviane HOOS / Dr. Bertrand RAYNAL
Secretary: 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 2010):

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

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

3) Fluorescence spectroscopy: Quantamaster QM4CW fluorimeter (Photon Technology International), with polarisation and time-resolved fluorescence modules ; KinetAsyst SF61-DX2 “stopped flow” instrument (TgK Scientific), with a polarisation module.

4) Infrared spectroscopy: FTIR 6100 spectrometer (Jasco), with an attenuated total reflectance accessory.

5) Light scattering: DynaPro MS800 and DynaPro PlateReader dynamic light scattering instruments (Wyatt) ; TDAmax light scattering-intrinsic viscosity-refractive index triple detector array (Malvern), coupled to a size exclusion chromatography system.

6) Microcalorimetry: ITC200, VP-ITC & MCS-ITC isothermal titration calorimeters ; VP-DSC differential scanning calorimeter, with a Pressure Perturbation Calorimetry accessory (PPC) (GE Healthcare).

7) Surface plasmon resonance: Biacore 2000 and Biacore X100 biosensors (GE Healthcare), 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 the following 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 17 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 22 laboratories from other institutions (french or foreign), leading to 43 peer-reviewed publications.

Below is a small sample of the PFBMI’s scientific collaborations since 2007:

1) Physico-chemical characterization of the properties of prolactin receptor antagonists (J. Biol. Chem (2007) 282, 33118-33131; J. Biol. Chem (2010) 285, 8422-8433; J. Mol. Biol. (2010) 404, 112-126)

2) Study of the conformational changes induced by calcium in the RTX subdomain of Bordetella pertussis adenylate cyclase toxin (J. Biol. Chem (2009) 284, 1781-1789; Biochemistry (2010) 49, 318-328; Biophys. J. (2010)99, 3744-3753)

3) Elucidation of the structural basis of the biological properties of the multimodular erythrocyte membrane protein 1 of Plasmodium falciparum (PfEMP1) (PNAS (2010) 107, 4884-4889; Mol. Biochem. Parasitol. (2010) 170, 84-92 and 173, 115-122)

4) 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)

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

6) Characterization of the specificity of the murine receptor FcγRIV towards immunoglobulins E and G (J. Clin. Invest. (2008) 118, 3738-3750; Blood (2009) 113, 3716-3725)

7) Physico-chemical characterization of the phosphorylation-dependent molecular switch of GarA, an FHA-domain containing protein of Mycobacterium tuberculosis (FEBS Lett. (2009) 583, 301-307)

8) Study of the mimicry of Shigella flexneri surface oligosaccharides by peptides (J. Mol. Biol. (2009) 388, 839-850)

9) Structure-function study of the recognition between small G proteins of the Arf family and thir JIP effectors (EMBO Journal (2009)28, 2835-2845)

10) Development of tandem proteins allowing to trap efficiently polyubiquitylated proteins (EMBO Rep (2009) 10, 1250-1258)

11) Study of the coupling between the dimerization of the NFkB modulator protein NEMO and its ubiquitin binding (J. Mol. Biol. (2010) 395, 89-104)

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



van Agthoven .J, Zhang C., Tallet E., Raynal B., Hoos S., Baron B., England P., Goffin V. & Broutin I. (2010)Structural characterization of the stem-stem dimerization interface between prolactin receptor chains complexed with the natural hormone.Journal of Molecular Biology, 404, 112-126

Srivastava A., Gangnard S., Round A., Dechavanne S., Juillerat A., Raynal B., Faure G., Baron B., Ramboarina S., Singh S.K., Belrhali H., England P., Lewit-Bentley A., Scherf A., Bentley G.A. & Gamain B. (2010)“Full-length extracellular region of the var2CSA variant of PfEMP1 is required for specific, high-affinity binding to CSA”Proceedings of the National Academy of Sciences of the USA (PNAS), 107, 4884-4889

England P., Wehenkel A., Martins S., Hoos S., André-Leroux G., Villarino A. & Alzari P.M. (2009)
"The FHA-containing protein GarA acts as a phosphorylation-dependent molecular switch in mycobacterial signaling" FEBS Letters, 583, 301-307

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 Chemistry 283, 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 Chemistry 283, 33551-33560

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Activity Reports 2010 - Institut Pasteur
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