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Adhesion Protein Protocols pp 251–271Cite as

Purification of Arp2/3 Complex from Saccharomyces cerevisiae

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1046))

Abstract

Much of the cellular control over actin dynamics comes through regulation of actin filament initiation. At the molecular level, this is accomplished through a collection of cellular protein machines, called actin nucleation factors, which position actin monomers to initiate a new actin filament. The Arp2/3 complex is a principal actin nucleation factor used throughout the eukaryotic family tree. The budding yeast Saccharomyces cerevisiae has proven to be not only an excellent genetic platform for the study of the Arp2/3 complex, but also an excellent source for the purification of endogenous Arp2/3 complex. Here we describe a protocol for the preparation of endogenous Arp2/3 complex from wild type Saccharomyces cerevisiae. This protocol produces material suitable for biochemical study and yields milligram quantities of purified Arp2/3 complex.

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Authors and Affiliations

  1. Department of Biophysics, UT Southwestern Medical Center and Howard Hughes Medical Institute, Dallas, TX, USA

    Lynda K. Doolittle, Michael K. Rosen & Shae B. Padrick

Authors
  1. Lynda K. Doolittle
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  2. Michael K. Rosen
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  3. Shae B. Padrick
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Editor information

Editors and Affiliations

  1. , Department of Oncology, University of Oxford, Roosevelt Drive, Oxford, OX3 7DQ, United Kingdom

    Amanda S. Coutts

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Doolittle, L.K., Rosen, M.K., Padrick, S.B. (2013). Purification of Arp2/3 Complex from Saccharomyces cerevisiae . In: Coutts, A. (eds) Adhesion Protein Protocols. Methods in Molecular Biology, vol 1046. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-538-5_15

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  • DOI: https://doi.org/10.1007/978-1-62703-538-5_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-537-8

  • Online ISBN: 978-1-62703-538-5

  • eBook Packages: Springer Protocols

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