Purification of Arp2/3 complex from saccharomyces cerevisiae

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

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages251-271
Number of pages21
Volume1046
ISBN (Print)9781627035378
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1046
ISSN (Print)10643745

Keywords

  • Actin nucleation factor
  • Arp2/3 complex
  • Biochemical purification
  • Detailed protocol
  • Endogenous source
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Doolittle, L. K., Rosen, M. K., & Padrick, S. B. (2013). Purification of Arp2/3 complex from saccharomyces cerevisiae. In Methods in Molecular Biology (Vol. 1046, pp. 251-271). (Methods in Molecular Biology; Vol. 1046). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-538-5_15