Institut de Chimie & Biologie des Membranes & des Nano-objets • Bordeaux

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Christian Griesinger, November 24th, 11am, IECB R. Dos santos Morais, 30/11/2017, 4 PM, ENSCBP Takahiro Muraoka, 16/11/2017, 11am, ENSCBP Vladimir Torbeev, 9/11/2017, 2pm, ENSCBP M. D. Smith & T.Constantieux, 3/11/2017, 2h30 pm, IECB James Nowick, 15 september 2017, 11am, IECB Emmanuelle Thinon, 28 June 2017, 11 am, IECB Caroline Tokarski, 15 June 2017, 4pm, ENSCBP Christian Salesse, 8 June 2017, 4pm, ENSCBP Patrice Rey, 01 June 2017, 2h30 pm, ENSCBP Christina Sizun, 18 May 2017, 4 pm, ENSCBP Marisela Velez, 5 May 2017, 11am, IECB Iqbal Choudhary, 27 April 2017, 4 pm, ENSCBP Vincent Aucagne, 21 April 2017, 11 am, IECB Sophie Zinn-Justin, 14 April 2017, 11 am, IECB Cécile Feuillie, 16 February 2017, 4 PM, ENSCBP Félix M. Goñi, 8 december 2016, 4 pm, ENSCBP Félix M. Goñi, 17 november 2016, 4 pm, ENSCBP Carl Creutz, 20 october 2016, 4 pm, ENSCBP Diego Romero, 30 september 2016, 11 am, IECB A. Ciaccafava, 8 september 2016, 14h, ENSCBP A. Ramamoorthy, 16 June 2016, 16h, ENSCBP Alexandre de Brevern, 2 june 2016, ENSCBP Isabelle Landrieu, 26 May, 16h, ENSCBP Frances Sepavoric, 12 May, 16h, ENSCBP Thomas Pradeu, 7 April 2016, 9h, ENSCBP Françoise Argoul, 3 march 2016, 16h, ENSCBP Corinne Loutelier-Bourhis, 16/12/2015, 16h Aristotelis XENAKIS, 3 december 2015, 16h Fabian Kiessling, 26 november 2015, 11h Michaël Molinari, 20 november 2015, 11h Dipankar Das Sarma, 28 October 2015, 14h. Olivier Donard, 22 october 2015, 16h, ENSCBP E. Morvan & A.Grelard, 17/12/2015, 16h, ENSCBP Christophe Cullin, 10 september 2015, 14 h Brigitte Lindet, 18 June 2015, 16h, ENSCBP(B) Marion Decossas, 4 June 2015, 16h, ENSCBP(B) Pascale Schellenberger, 21 Mai 2015, 16h F. Leal-Calderon, 7 May 2015, 16h, ENSCBP(B) Ibrahim Abdulhalim, 9 April 2015, 14h, ENSCBP Manon Carré, 26 March 2015, 14h, ENSCBP(B) C. Bure & JM Schmitter, 19 March 2015, 16h T. Ogata & H. Ihara, 17 March 2015, 11h, IECB Banafshe Larijani, 12 March 2015, 16h, ENSCBP
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Christian Griesinger, November 24th... R. Dos santos Morais, 30/11/2017, 4... Takahiro Muraoka, 16/11/2017, 11am,... Vladimir Torbeev, 9/11/2017, 2pm, E... M. D. Smith & T.Constantieux, 3/11/... James Nowick, 15 september 2017, 11... Emmanuelle Thinon, 28 June 2017, 11... Caroline Tokarski, 15 June 2017, 4p... Christian Salesse, 8 June 2017, 4pm... Patrice Rey, 01 June 2017, 2h30 pm,... Christina Sizun, 18 May 2017, 4 pm,... Marisela Velez, 5 May 2017, 11am, I... Iqbal Choudhary, 27 April 2017, 4 p... Vincent Aucagne, 21 April 2017, 11 ... Sophie Zinn-Justin, 14 April 2017, ... Cécile Feuillie, 16 February 2017,... Félix M. Goñi, 8 december 2016, 4... Félix M. Goñi, 17 november 2016, ... Carl Creutz, 20 october 2016, 4 pm,... Diego Romero, 30 september 2016, 11... A. Ciaccafava, 8 september 2016, 14... A. Ramamoorthy, 16 June 2016, 16h,... Alexandre de Brevern, 2 june 2016, ... Isabelle Landrieu, 26 May, 16h, ENS... Frances Sepavoric, 12 May, 16h, ENS... Thomas Pradeu, 7 April 2016, 9h, EN... Françoise Argoul, 3 march 2016, 1... Corinne Loutelier-Bourhis, 16/12/20... Aristotelis XENAKIS, 3 december 201... Fabian Kiessling, 26 november 2015,... Michaël Molinari, 20 november 2015... Dipankar Das Sarma, 28 October 2015... Olivier Donard, 22 october 2015, 16... E. Morvan & A.Grelard, 17/12/2015, ... Christophe Cullin, 10 september 201... Brigitte Lindet, 18 June 2015, 16h,... Marion Decossas, 4 June 2015, 16h, ... Pascale Schellenberger, 21 Mai 2015... F. Leal-Calderon, 7 May 2015, 16h, ... Ibrahim Abdulhalim, 9 April 2015, 1... Manon Carré, 26 March 2015, 14h, ... C. Bure & JM Schmitter, 19 March 20... T. Ogata & H. Ihara, 17 March 2015,... Banafshe Larijani, 12 March 2015, 1...
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Patricia DULOR Anthony BOUTER Olivier LAMBERT Jochen LANG Pascale SUBRA-PATERNAULT Maria URDACI Marie-Christine DURRIEU Xavier SANTARELLI Gilles GUICHARD Yann FERRAND Isabelle BESTEL Juan ELEZGARAY Antoine LOQUET Sophie LECOMTE Reiko ODA Marc BONNEU

R. Dos santos Morais, 30/11/2017, 4 PM, ENSCBP

The seminar will take place, the 30th of November 2017, at 4pm, at the  ENSCBP amphitheater, Building B,  allée Geoffroy Saint-Hilaire, Pessac.

 

Prior to the seminar, NanoTemper Technologies will introduce MST and nanoDSF as the technique have been used to conduct the following research.

 

Dystrophin-membrane interactions: evidence of structural changes upon binding to phospholipids 

 

Dr. Raphael Dos Santos Morais,

Université de Rennes 1, F-35043 Rennes, France.

Institut de Génétique et Développement de Rennes, CNRS UMR 6290, F-35043 Rennes, France. Laboratoire Léon-Brillouin, UMR 12 CEA-CNRS, Université Paris-Saclay, CEA-Saclay, F-91191Gif-sur-Yvette CEDEX, France.

SWING Beamline, Synchrotron SOLEIL, L'Orme des Merisiers, BP 48, Saint-Aubin, F-91192 Gif-sur-Yvette, France.

 

Abstract

 

Dystrophin1 is a peripheral membrane protein supporting the plasma membrane of muscle cells. Gene mutations lead either to a total deficit of dystrophin or to the presence of dystrophin in truncated forms and are responsible of Duchenne or Becker muscular dystrophy respectively. Exhaustive knowledge of dystrophin/membrane interactions is needed to contribute to gene therapy strategies devoted to DMD and BMD, aiming to design a minimal functional protein. Most part of the dystrophin consists in a central domain, made of 24 coiled-coil repeats (R1 to R24), whose the 3D-structure is not accessible by classical high resolution methods2.

 

First, protein/lipid interactions of two lipid-binding subdomains3–6, R1-3 and R11-15, were characterized with phospholipid-based bicelles as a membrane mimic7. We notably used Microscale thermophoresis8 (MST, NanoTemper Techonologies, Figure 1A) to highlight protein/bicelle interactions and to determine a dissociation constant of the complexes. To our knowledge, it is the first time that bicelles are used in MST experiments and we do believe that this approach could be extended to others protein/lipid systems. Then; using small-angle neutron scattering (SANS), we specifically probed (i) the solution structure of R1-3 and R11-15 and (ii) the solution structure of R1-3 and R11-15 in the presence of either zwitterionic (ZB) or anionic bicelles (AB), in order to probe the role of the electrostatic properties of the membrane.

 

Our results highlight that, when bound to ZB, no important conformational modification of the protein fragments are detected. On the contrary, when the protein fragments are bound to AB, especially for R1-3, SANS data demonstrate large modifications of their 3D-structures. The R1-3/AB complex was further analyzed by classical coarse-grained molecular dynamic simulations (CG-MD) and interactive CG-MD9 (CG-IMD).The final models proposed for R1-3 bound to membrane lipids are totally in adequacy with SANS data as well as with the mapping of protein/lipid interaction obtained by coupling click-chemistry with mass spectrometry (Figure 1B). We hypothesize that the membrane anchoring that likely occurs during the contraction/elongation process of muscles could be ensured by the opening of the first spectrin-like repeat of R1-3. Understanding these structural changes may help for the design of rationalized efficient shortened dystrophins devoted to gene therapy. Last but not least, we expect that our approaches can be extended to other peripheral and even integral membrane proteins.

 

 

 Bibliography

 

 (1) Le Rumeur, E.; Winder, S. J.; Hubert, J.-F. Dystrophin: More than Just the Sum of Its Parts. Biochim. Biophys. Acta BBA - Proteins Proteomics 2010, 1804 (9), 1713–1722.

(2) Molza, A.-E.; Férey, N.; Czjzek, M.; Le Rumeur, E.; Hubert, J.-F.; Tek, A.; Laurent, B.; Baaden, M.; Delalande, O. Innovative Interactive Flexible Docking Method for Multi-Scale Reconstruction Elucidates Dystrophin Molecular Assembly. Faraday Discuss. 2014, 169, 45–62.

(3) Legardinier, S.; Hubert, J.-F.; Le Bihan, O.; Tascon, C.; Rocher, C.; Raguénès-Nicol, C.; Bondon, A.; Hardy, S.; Le Rumeur, E. Sub-Domains of the Dystrophin Rod Domain Display Contrasting Lipid-Binding and Stability Properties. Biochim. Biophys. Acta 2008, 1784 (4), 672–682.

(4) Legardinier, S.; Raguénès-Nicol, C.; Tascon, C.; Rocher, C.; Hardy, S.; Hubert, J.-F.; Le Rumeur, E. Mapping of the Lipid-Binding and Stability Properties of the Central Rod Domain of Human Dystrophin. J. Mol. Biol. 2009, 389 (3), 546–558.

(5) Vié, V.; Legardinier, S.; Chieze, L.; Le Bihan, O.; Qin, Y.; Sarkis, J.; Hubert, J.-F.; Renault, A.; Desbat, B.; Le Rumeur, E. Specific Anchoring Modes of Two Distinct Dystrophin Rod Sub-Domains Interacting in Phospholipid Langmuir Films Studied by Atomic Force Microscopy and PM-IRRAS. Biochim. Biophys. Acta 2010, 1798 (8), 1503–1511.

(6) Sarkis, J.; Hubert, J.-F.; Legrand, B.; Robert, E.; Chéron, A.; Jardin, J.; Hitti, E.; Le Rumeur, E.; Vié, V. Spectrin-like Repeats 11-15 of Human Dystrophin Show Adaptations to a Lipidic Environment. J. Biol. Chem. 2011, 286 (35), 30481–30491.

(7) Dos Santos Morais, R.; Delalande, O.; Pérez, J.; Mouret, L.; Bondon, A.; Martel, A.; Appavou, M.-S.; Le Rumeur, E.; Hubert, J.-F.; Combet, S. Contrast-Matched Isotropic Bicelles: A Versatile Tool to Specifically Probe the Solution Structure of Peripheral Membrane Proteins Using SANS. Langmuir 2017, 33 (26), 6572–6580.

(8) Jerabek-Willemsen, M., André, T., Wanner, R., Roth, H. M., Duhr, S., Baaske, P., and Breitsprecher,D. (2014) MicroScale Thermophoresis: Interaction analysis and beyond. Journal of Molecular Structure

(9) Delalande, O.; Férey, N.; Grasseau, G.; Baaden, M. Complex Molecular Assemblies at Hand via Interactive Simulations. J. Comput. Chem.2009, 30 (15), 2375–2387.

 

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