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Updated: Sep 3, 2026

Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin
Published on: March 28, 2008
In Vitro Analysis of Profilin 1 Binding to Recombinant Actin
Leticia Villadangos1, David Escot1, Juan M Serrador2
1Interactions with the Environment Program, Development and Function of the Immune System Unit, Centro de Biología Molecular Severo Ochoa (CBMSO), Consejo Superior de Investigaciones Científicas (CSIC)-Universidad Autónoma de Madrid (UAM), Madrid, Spain.
Abstract:
Profilin 1 (PFN1) is a key actin-binding protein (ABP) involved in cytoskeletal dynamics, regulating actin polymerization and filament remodeling. While native actin has been extensively studied, recombinant actin produced through biotechnological approaches remains less explored due to solubility and folding challenges. Previously, we assessed the effects of S-nitrosylation on actin binding, providing insights into post-translational modifications that regulate actin function. In the present study, we provide a comprehensive protocol to produce recombinant β-actin using an in vitro transcription-translation system supplemented with chaperonin CCT to analyze its interaction with PFN1. Our findings highlight the utility of recombinant actin for studying ABPs and their regulatory mechanisms, offering a cost-effective alternative for structural and functional analyses.
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