Protocol

Adhesion Protein Protocols

Volume 1046 of the series Methods in Molecular Biology pp 251-271

Date:

Purification of Arp2/3 Complex from Saccharomyces cerevisiae

  • Lynda K. DoolittleAffiliated withDepartment of Biophysics, UT Southwestern Medical Center and Howard Hughes Medical Institute
  • , Michael K. RosenAffiliated withDepartment of Biophysics, UT Southwestern Medical Center and Howard Hughes Medical Institute
  • , Shae B. PadrickAffiliated withDepartment of Biophysics, UT Southwestern Medical Center and Howard Hughes Medical Institute

* Final gross prices may vary according to local VAT.

Get Access

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.

Key words

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