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Christophe Bucher, Thursday July 18... Mélanie Berbon, 11 juillet 2019, 1... Hendrik W. Van Veen, July 11th, 201... Sébastien Ulrich, July 4th, 2019 ... Frédéric Brossard, 1st of July, 1... Prof. Solange Landreville, 19th of ... Prof. Hiroshi Yabu, 27th of May, 11... Amitabha Chattopadhyay, 22 May, 11 ... Prof. Hiroaki Suga, 10/05/2017, 11 ... INTERNAL CBMN SEMINAR: PHD student ... Jean Duhamel, 4 april 2019, 11am, I... Internal CBMN seminar 7 March 2019... Sandrine VIllette, 7 february 2019,... Internal CBMN Seminar : newcomers ... Internal CBMN Seminar: A. Simon, 6 ... Marisela Velez, 28 november 2018, 2... Marc Bramkamp, 16 november 2018, 2p... Kazushi Kinbara, 12 november 2018, ... Internal CBMN Seminar: J. Lang, 8 N... Internal CBMN Seminars: A. Baudin, ... Plateforme production de protéine... Patrick Trouillas, 21 June 2018, 2p... Lucie Khemtemourian, 21 June 2018, ... Jonathan Faherty, 7th June, 11 AM, ... Yann Fichou, 3 May 2018, 2pm, IECB Internal CBMN seminars: A. Ciaccafa... Alain Roussel, 29/03/2018, 4 pm, EN... Simon Poly, 22 march 2018, 2pm, ENS... J.Crassous, M. Sallé & A. Martinez... O.Reinaud & V. Artero, 7/12/2017, 1... R. Dos santos Morais, 30/11/2017, 4... Christian Griesinger, November 24th... 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 Caroline TOKARSKI

Jean Duhamel, 4 april 2019, 11am, IECB

The seminar will take place, the 4th of April, at 11 am, in the IECB amphitheatre, 2 rue Robert Escarpit Pessac.

 

Helices in biological and synthetic macromolecules probed by time-resolved fluorescence

 

Jean Duhamel

Institute for Polymer Research, Waterloo Institute for Nanotechnology, Department of Chemistry, University of Waterloo, Canada

 

Abstract

 

In 1967, Stryer and Haugland demonstrated that fluorescence resonance energy transfer (FRET) between a naphthyl donor and a dansyl acceptor could be applied to probe the distance between the two ends of a poly(L-proline) type II trans helix that had been end-labeled with these dyes. This study defined FRET as a Spectroscopic Ruler which explains the supremacy of FRET to this date among other fluorescence techniques when seeking structural information about macromolecules. Yet time-resolved fluorescence anisotropy (TRFA) and pyrene excimer fluorescence (PEF) are becoming increasingly used to characterize the helical nature of well and poorly defined macromolecules.

 

This presentation will illustrate the applicability of these methods based on TRFA and PEF. In particular, the ability of TRFA to determine the hydrodynamic volume of rigid macromolecular objects was applied to demonstrate that well-defined oligoquinoline foldamers labeled at one end with an oligo(phenylene vinylene) dye retained a helical conformation in chloroform. By contrast, PEF can be applied to probe the helical nature of poorly defined macromolecules, i.e. polydisperse macromolecules of different lengths but of same chemical composition. PEF does this by determining the dimension of the restricted volume where an excimer can be formed from the encounter between an excited and a ground-state pyrene covalently attached to a macromolecule. Analysis of the fluorescence decays acquired with pyrene-labeled macromolecules (PyLMs) yields the dimension of the restricted volume, also referred to as a blob from the knowledge of the pyrene content of the PyLM. Molecular Mechanics Optimizations (MMOs) can then be employed to assess whether the size of a blob is compatible with a helical conformation. This feature has been applied to demonstrate that poly(L-glutamic acid) in DMF1,2 and amylose in DMSO3 both adopt helical conformations. An additional feature of handling PEF with the FBM is that it provides information about interhelical distances between helices constituting the building blocks of a macromolecule. This was used to determine the interhelical distance between the strands of oligo(L-glutanic acid)4 and oligosaccharide helices constituting a poly(L-glutamic acid) arborescent polymer and amylopectin.

 

 

Recent Publications

  1. Duhamel, J.; Kanagalingam, S.; O’Brien, T.; Ingratta, M. Side-Chain Dynamics of an Helical Polypeptide Monitored by Fluorescence. J. Am. Chem. Soc.2003, 125, 12810-12822.

  2. Casier, R.; Duhamel, J. Pyrene Excimer Fluorescence as a Direct and Easy Experimental Means to Characterize the Length Scale and Dynamics of Polypeptide Foldons. Macromolecules 2018, 51, 3450-3457.

  3. Li, L.; Duhamel, J. Conformation of Pyrene-Labeled Amylose in DMSO Characterized with the Fluorescence Blob Model. Macromolecules 2016, 49, 7965-7974.

  4. Hall, T.; Whitton, G.; Casier, R.; Gauthier, M.; Duhamel, J. Arborescent Poly(L-glutamic acid)s as Standards to Study the Dense Interior of Polypeptide Mesoglobules by Pyrene Excimer Fluorescence. Macromolecules2018, 51, 7914-7923.

 

 

 

Biography

After being trained as a chemical engineer in France at the Ecole Nationale Supérieure des Industries Chimiques, Jean Duhamel obtained his PhD in 1989 under the supervision of Dr. André at the Institut National Polytechnique de Lorraine (France). Following a first post-doc at the U. of Toronto (Canada) with Prof. Winnik and a second one at the U. of Pennsylvania (USA) with Prof. Lu, he joined the U. of Waterloo in 1996. Over the past 20+ years, work from his research group has aimed to establish the versatility of pyrene excimer fluorescence or PEF to probe the dynamics, conformation, and interactions of any macromolecule in solution.He has published over 100 refereed papers to date with a third in Macromolecules.

 

E-mail: jduhamel@uwaterloo.ca

 

Contact: Victor Maurizot (v.maurizot@cbmn.u-bordeaux.fr)

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