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

Christine Ebel; Catherine Birck

doi:10.1007/978-1-0716-1126-5_9

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

Methods Mol Biol. 2021:2247:155-171. doi: 10.1007/978-1-0716-1126-5_9.

Authors

Christine Ebel¹, Catherine Birck²

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@ibs.fr.
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@igbmc.fr.

PMID: 33301117
DOI: 10.1007/978-1-0716-1126-5_9

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.

Keywords: Analytical ultracentrifugation; Hetero-association; Protein heterogeneity; Protein interaction; Sedimentation coefficient; Sedimentation velocity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Algorithms
Biophysical Phenomena*
Biosensing Techniques*
Data Analysis
Models, Theoretical
Protein Binding
Proteins / chemistry*
Temperature
Ultracentrifugation / methods

Substances

Proteins