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NeuroTimone Interactome Platform "PINT" - Analytical Ultracentrifugation Platform

Our laboratory (Biophysics, CNRS then Inserm Unit) has been using Analytical Ultra-Centrifugation (UCA) for more than 25 years to study proteins and their self-associative processes. We have analyzed the formation of ring-shaped polymers of tubulin after the proteolytic cleavage of the C-terminal domains of both alpha and beta tubulin chains by subtilisin (Peyrot et al. , 1990;. Peyrot et al. , 1991). The formation of tubulin polymer induced by the anti-cancer drug cryptophycin 52 (Barbier et al., 2001) has also been extensively characterized. We have applied UCA to the interaction between tau and microtubules (Devred et al., 2004, Amniai et al 2009, Gigant et al., 2014) and also, associated with microcalorimetry techniques, on Stathmin-tubulin interactions (Devred et al. , 2008, Barbier et al., 2010). The oligomerization states of HSP90 and its multiprotein complexes (Garnier et al., 2002, Moullintraffort et al., 2010, Lepvrier et al., 2015); those of Nipah virus (NiV) viral proteins (Blocquel et al., 2013; Bertrandi et al . , 2015) as well as the interaction between cytochrome C3 type I and II (Pieulle et al. , 2005) were addressed by UCA.

Analytical UltraCentrifugation (UCA) studies the behavior of native and aqueous solution macromolecules subjected to centrifugal force. With the help of a detection system that allows us to measure the concentration of macromolecules as a function of the centrifugation radius, we can carry out two types of experiments that will lead to obtaining additional information:

The sedimentation rate allows us to obtain the sedimentation coefficient, the shape, the associative state (monomer, dimer, trimer,...) of a macromolecule. It can also highlight the formation of a complex and allow the determination of its affinity constant.

The sedimentation equilibrium allows to obtain the mass, the associative state (monomer, dimer, trimer,...) and to highlight the formation of a macromolecular complex.

MEMBERS

Regular / Staff members

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Diane
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ALLEGRO
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Assistante Ingénieur
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Pascale
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BARBIER
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Directeur de l'Unité - MCU
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Vincent
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PEYROT
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Professeur
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RECENT PUBLICATIONS

  • Beltrandi, M. Blocquel, D. Eralesa,J. Barbier P. Cavalli A. Longhi, S. Insights into the coiled-coil organization of the Hendra virus phosphoprotein from combined biochemical and SAXS studies. (2015), Virology 477:42-55.
  • Blocquel D., Beltrandi M., Erales J. Barbier P, Longhi S. Biochemical and structural studies of the oligomerization domain of the Nipah virus phosphoprotein: evidence for an elongated coiled-coil homotrimer. Virology. (2013) 446(1–2): 162-72.
  • Barbier P, Dorleans A, Devred F, Sanz L, Allegro D, Alfonso C, Knossow M, Peyrot V, Andreu JM. Stahtmine and interfacial microtubule inhibitors recognize a naturally curved conformation of tubulin dimers. Journal of biological Chemistry, (2010) 285(41):31672-81.
  • Devred F, Barbier P, Lafitte D, Landrieu I, Lippens G, Peyrot V. Microtubule and MAPs: thermodynamics of complex formation by AUC, ITC, fluorescence, and NMR. Methods in Cellular Biology (2010);95:449-80.