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Macromolecular interactions in vitro, comparing classical and novel approaches

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Title: Macromolecular interactions in vitro, comparing classical and novel approaches
Authors: Velours, Christophe, Aumont-Nicaise, Magali, Uebel, Stephan, England, Patrick, Velazquez-Campoy, Adrian, Stroebel, David, Bec, Guillaume, Soule, Pierre, Quétard, Christophe, Ebel, Christine, Roussel, Alain, Charbonnier, Jean-Baptiste, Varela, Paloma Fernández
Contributors: Agence Nationale de la Recherche (France), univOAK, Archive ouverte, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Microbiologie Fondamentale et Pathogénicité (MFP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Max-Planck-Institut für Biochemie = Max Planck Institute of Biochemistry (MPIB), Max-Planck-Gesellschaft, Biophysique Moléculaire (plateforme) - Molecular Biophysics (platform), Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS), Universidad de Zaragoza = University of Zaragoza Saragossa University = Université de Saragosse, Instituto de Investigación Sanitaria de Aragón Zaragoza (IIS Aragón), Centro de Investigación Biomédica en Red en el Área temática de Enfermedades Hepáticas y Digestivas (CIBERehd), Liver Unit, Clínica Universitaria, CIBER-EHD, Fundación Agencia Aragonesa para la Investigación y el Desarrollo (ARAID), Institut de biologie de l'ENS Paris (IBENS), Département de Biologie - ENS-PSL (IBENS), École normale supérieure - Paris (ENS-PSL), Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Architecture et Réactivité de l'ARN (ARN), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), NanoTemper Technologies GmbH München, ForteBio-Sartorius Dourdan, Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), JBC is supported by ARC program (SLS220120605310), French Infrastructure for Integrated Structural Biology (FRISBI) ANR-10-INSB-05, ANR-18-CE44-0008, ANR-20-CE11-0026, INCA 2016-1-PL BIO-11 and INCA SLX4 INCA 2016-159., ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), ANR-18-CE44-0008,DNAPAR18,Etude de l'ADP-ribosylation d'ADN et de son rôle dans la réponse aux dommages à l'ADN(2018), ANR-20-CE11-0026,BreakDance,Caractérisation de la chorégraphie orchestrée par l'hétérodimère Ku sur les cassures double-brin de l'ADN(2020)
Source: Digital.CSIC. Repositorio Institucional del CSIC
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Publisher Information: Springer Science and Business Media LLC, 2021.
Publication Year: 2021
Subject Terms: 0301 basic medicine, 0303 health sciences, Macromolecular Substances, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Proteins, DNA, Calorimetry, Ligands, Artificial binders, 3. Good health, 03 medical and health sciences, Artifcial binders, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Humans, Double-stranded DNA breaks repair factors, Molecular Biology, Molecular scale biophysics, Macromolecular interactions
Description: 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 ( https://mosbio.sciencesconf.org/ ). 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.
Document Type: Article
File Description: application/pdf
Language: English
ISSN: 1432-1017
0175-7571
DOI: 10.1007/s00249-021-01517-5
Access URL: https://hal.archives-ouvertes.fr/hal-03224764/file/islandora_117196.pdf
https://pubmed.ncbi.nlm.nih.gov/33792745
http://hdl.handle.net/10261/253612
https://pubmed.ncbi.nlm.nih.gov/33792745/
https://europepmc.org/article/MED/33792745
https://digital.csic.es/handle/10261/253612
https://link.springer.com/article/10.1007/s00249-021-01517-5
Rights: Springer TDM
Accession Number: edsair.doi.dedup.....bc11bcc7f6e9550ab8b8f56d6283fd82
Database: OpenAIRE
Description
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 ( https://mosbio.sciencesconf.org/ ). 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.
ISSN:14321017
01757571
DOI:10.1007/s00249-021-01517-5