title: Purification and architecture of the ubiquitous Wave complex.
abstract: The Wave proteins are major activators of the Arp2/3 complex. The ubiquitous Wave-2 is required for actin polymerization at the leading edge of migrating cells. Here we purify Wave-2 from HeLa cells. Five proteins, Sra, Nap, Wave-2, Abi, and Hspc, are copurified, indicating that they form a tight complex. These proteins are only present in the complexed form, with the exception of Hspc, which displays a free pool. We reconstitute the Wave-2 complex by cotranslating in vitro the five subunits and use this system together with specific immunoprecipitations to study the molecular architecture of the complex. The complex is organized around a core of Nap and Abi. Sra is a peripheral subunit recruited on the Nap side, whereas the Wave and Hspc subunits are recruited on the Abi side of the core.
authors.affiliation: Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
authors.forename: Alexis
authors.initials: A
authors.lastname: Gautreau
authors.forename: Hsin-yi H
authors.initials: HY
authors.lastname: Ho
authors.forename: Jiaxu
authors.initials: J
authors.lastname: Li
authors.forename: Hanno
authors.initials: H
authors.lastname: Steen
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
authors.forename: Marc W
authors.initials: MW
authors.lastname: Kirschner
mesh_terms: D000595:Amino Acid Sequence; D002499:Centrifugation, Density Gradient; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D008840:Microfilament Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D021122:Protein Subunits / Q000737:chemistry; D051300:Wiskott-Aldrich Syndrome Protein Family
chemical_list: D018076:DNA, Complementary; D008840:Microfilament Proteins; D010446:Peptide Fragments; D021122:Protein Subunits; C120152:WASF2 protein, human; D051300:Wiskott-Aldrich Syndrome Protein Family