Unit

UNIT 13.6 Functional Characterization of Proteins Regulating Actin Assembly

  1. Maud Hertzog1,
  2. Marie-France Carlier2

Published Online: 1 APR 2005

DOI: 10.1002/0471143030.cb1306s26

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Hertzog, M. and Carlier, M.-F. 2005. Functional Characterization of Proteins Regulating Actin Assembly. Current Protocols in Cell Biology. 26:13.6:13.6.1–13.6.23.

Author Information

  1. 1

    Istituto FIRC di Oncologia Molecolare Fondazione Italiana per la Ricerca sul Cancro, Milano, Italy

  2. 2

    Laboratoire d'Enzymologie et Biochimie Structurale Centre National de la Recherche Scientifique, Gif-sur-Yvette, France

Publication History

  1. Published Online: 1 APR 2005
  2. Published Print: MAR 2005

Abstract

A very large, ever-increasing repertoire of actin-binding proteins regulates the assembly dynamics and the spatial organization of actin filaments, thus orchestrating the motile behavior of the cell. The authors describe a series of biochemical functional assays that allow one to characterize the function of a putative actin-binding protein in actin filament dynamics. These tests allow the characterization of three types of actin-binding proteins: G-actin-sequestering proteins, profilin-like proteins, and barbed-end capping proteins. Biochemical tests include the use of sedimentation of actin filaments, polymerization assays at the barbed or pointed end of actin filaments derived from fluorescently labeled actin, thermodynamic measurements of actin assembly at steady state and during turnover of actin filaments, measurements of nucleotide exchange on G-actin, and the use of the intrinsic or extrinsic fluorescence of actin to measure direct binding of different protein ligands to G-actin.

Keywords:

  • actin;
  • polymerization;
  • actin-binding proteins;
  • ADF/cofilin;
  • profilin;
  • actin-sequestering β-thymosins;
  • capping proteins