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  Macromolecular interactions in vitro, comparing classical and novel approaches

Velours, C., Aumont-Nicaise, M., Uebel, S., England, P., Velazquez-Campoy, A., Stroebel, D., et al. (2021). Macromolecular interactions in vitro, comparing classical and novel approaches. European Biophysics Journal, 50, 313-330. doi:10.1007/s00249-021-01517-5.

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 Creators:
Velours, Christophe1, Author
Aumont-Nicaise, Magali1, Author
Uebel, Stephan2, Author              
England, Patrick1, Author
Velazquez-Campoy, Adrian1, Author
Stroebel, David1, Author
Bec, Guillaume1, Author
Soule, Pierre1, Author
Quetard, Christophe1, Author
Ebel, Christine1, Author
Roussel, Alain1, Author
Charbonnier, Jean-Baptiste1, Author
Varela, Paloma Fernandez1, Author
Affiliations:
1external, ou_persistent22              
2Scientific Service Groups, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565170              

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Free keywords: Biophysics; Molecular scale biophysics; Macromolecular interactions; Artificial binders; Double-stranded DNA breaks repair factors;
 Abstract: Biophysical quantification of protein interactions is central to unveil the molecular mechanisms of cellular processes. Researchers can choose from a wide panel of biophysical methods that quantify molecular interactions in different ways, including both classical and more novel techniques. We report the outcome of an ARBRE-MOBIEU training school held in June 2019 in Gif-sur-Yvette, France (). Twenty European students benefited from a week's training with theoretical and practical sessions in six complementary approaches: (1) analytical ultracentrifugation with or without a fluorescence detector system (AUC-FDS), (2) isothermal titration calorimetry (ITC), (3) size exclusion chromatography coupled to multi-angle light scattering (SEC-MALS), (4) bio-layer interferometry (BLI), (5) microscale thermophoresis (MST) and, (6) switchSENSE. They implemented all these methods on two examples of macromolecular interactions with nanomolar affinity: first, a protein-protein interaction between an artificial alphaRep binder, and its target protein, also an alphaRep; second, a protein-DNA interaction between a DNA repair complex, Ku70/Ku80 (hereafter called Ku), and its cognate DNA ligand. We report the approaches used to analyze the two systems under study and thereby showcase application of each of the six techniques. The workshop provided students with improved understanding of the advantages and limitations of different methods, enabling future choices concerning approaches that are most relevant or informative for specific kinds of sample and interaction.

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Language(s): eng - English
 Dates: 2021
 Publication Status: Published in print
 Pages: 18
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Title: European Biophysics Journal
Source Genre: Journal
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Publ. Info: Berlin : Springer
Pages: - Volume / Issue: 50 Sequence Number: - Start / End Page: 313 - 330 Identifier: ISSN: 0175-7571
CoNE: https://pure.mpg.de/cone/journals/resource/954925487773_1