Biophysics of Macromolecules and their Interactions


  HEADDr ENGLAND Patrick / england@pasteur.fr
  MEMBERSHOOS Sylviane


  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.

Created in 2002, the Centre of Biophysics of Macromolecules and their Interactions (PFBMI) aims to group state-of-the-art equipment, adapted 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:

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 biosensor (Biacore).

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, biophysique@pasteur.fr . Instructions and down-loadable forms are available 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.

The 13 instruments and the know-how of the PFBMI have attracted more than 110 projects altogether, in association with 29 Research Units of the Pasteur Institute (from 9 out of 10 Scientific Departments) and 27 laboratories from other institutions (French or foreign), leading to 28 peer-reviewed publications.

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

* Elucidation of the role of several post-translational modifications in the tethering of protein HP1 to histone tails

* Characterization of the coupling between the trimerization of the rabies virus glycoprotein and its interaction with nerve growth factor receptor p75NTR

* Determination of the effect of natural pathogenic mutations of the NF-κB modulator protein NEMO on its stability and oligomerization properties

* Calorimetric characterization of the oligomeric structure of E. coli GMP kinase

* Optimization of the determination of the secondary structure of proteins by ATR infrared spectroscopy

* Federative Horizontal Program on Mycobacterium tuberculosis target discovery and drug design

england.jpg

figure 1: Etude de la dénaturation thermique d'une protéine suivie par dichroïsme circulaire dans l'UV lointain.
figure 2: Détecteur multiple Viscotek TDA302 couplé à un système de chromatographie d'exclusion stérique GPCmax (installé en novembre 2006)



  Web Site

More informations on our web site


  Publications

Publications 2006 of the unit on Pasteur's references database




Activity Reports 2006 - Institut Pasteur
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