Overview of current methods in sedimentation velocity and sedimentation equilibrium analytical ultracentrifugation

Huaying Zhao, Chad A Brautigam, Rodolfo Ghirlando, Peter Schuck

Research output: Contribution to journalArticle

102 Scopus citations

Abstract

Modern computational strategies have allowed for the direct modeling of the sedimentation process of heterogeneous mixtures, resulting in sedimentation velocity (SV) sizedistribution analyses with significantly improved detection limits and strongly enhanced resolution. These advances have transformed the practice of SV, rendering it the primary method of choice for most existing applications of analytical ultracentrifugation (AUC), such as the study of protein self- and hetero-association, the study of membrane proteins, and applications in biotechnology. New global multisignal modeling and mass conservation approaches in SV and sedimentation equilibrium (SE), in conjunction with the effective-particle framework for interpreting the sedimentation boundary structure of interacting systems, as well as tools for explicit modeling of the reaction/ diffusion/sedimentation equations to experimental data, have led to more robust and more powerful strategies for the study of reversible protein interactions and multiprotein complexes. Furthermore, modern mathematical modeling capabilities have allowed for a detailed description of many experimental aspects of the acquired data, thus enabling novel experimental opportunities, with important implications for both sample preparation and data acquisition. The goal of the current unit is to describe the current tools for the study of soluble proteins, detergent-solubilized membrane proteins and their interactions by SV and SE.

Original languageEnglish (US)
Article number20.12
JournalCurrent Protocols in Protein Science
Issue numberSUPPL.71
DOIs
StatePublished - Feb 1 2013

Keywords

  • Analytical ultracentrifugation
  • Chemical equilibria
  • Multiprotein complex
  • Protein hydrodynamics
  • Reversible interaction
  • Sedimentation equilibrium
  • Sedimentation velocity
  • Size-distribution

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry

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