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Sedimentation Velocity Methods for the Characterization of Protein Heterogeneity and Protein Affinity Interactions

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Multiprotein Complexes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2247))

Abstract

Sedimentation velocity analytical ultracentrifugation is a powerful and versatile tool for the characterization of proteins and macromolecular complexes in solution. The direct modeling of the sedimentation process using modern computational strategies allows among others to assess the homogeneity/heterogeneity state of protein samples and to characterize protein associations. In this chapter, we will provide theoretical backgrounds and protocols to analyze the size distribution of protein samples and to determine the affinity of protein–protein hetero-associations.

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Acknowledgments

The authors acknowledge the support and the use of resources of the French Infrastructure for Integrated Structural Biology FRISBI ANR-10-INBS-05 and of Instruct-ERIC: the platforms of the Grenoble Instruct-ERIC center (ISBG: UMS 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology (PSB) and the Strasbourg Instruct-ERIC center (Centre de Biologie Intégrative, CBI) within IGBMC (CNRS UMR 7104-Inserm U 1258-Université de Strasbourg). CE also acknowledges the support of the COST action CA 15126 MOBIEU and GRAL financed within the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003).

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Authors and Affiliations

  1. Institut de Biologie Structurale (IBS), Univ Grenoble Alpes, Commissariat à l’énergie atomique et aux énergies alternatives (CEA), Centre National de la Recherche Scientifique (CNRS), Grenoble, France

    Christine Ebel

  2. Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS), UMR 7104, Institut National de la Santé et de la Recherche Médicale (INSERM), U1258, Université de Strasbourg, Illkirch, France

    Catherine Birck

Authors
  1. Christine Ebel
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  2. Catherine Birck
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Corresponding authors

Correspondence to Christine Ebel or Catherine Birck .

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Editors and Affiliations

  1. Institut de Génétique et de Biologie Moléculaire et Cellulaire, Center for Integrated Biology, Integrated Structural Biology Department, Equipe labellisée Ligue Contre le Cancer, CNRS UMR 7104 - Inserm U 1258, University of Strasbourg, Illkirch, France

    Arnaud Poterszman

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Ebel, C., Birck, C. (2021). Sedimentation Velocity Methods for the Characterization of Protein Heterogeneity and Protein Affinity Interactions. In: Poterszman, A. (eds) Multiprotein Complexes. Methods in Molecular Biology, vol 2247. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1126-5_9

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  • DOI: https://doi.org/10.1007/978-1-0716-1126-5_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1125-8

  • Online ISBN: 978-1-0716-1126-5

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