TY - JOUR
T1 - Purification and architecture of the ubiquitous Wave complex
AU - Gautreau, Alexis
AU - Ho, Hsin-Yi Henry
AU - Li, Jiaxu
AU - Steen, Hanno
AU - Gygi, Steven P.
AU - Kirschner, Marc W.
PY - 2004/3/30
Y1 - 2004/3/30
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=1842428821&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=1842428821&partnerID=8YFLogxK
U2 - 10.1073/pnas.0400628101
DO - 10.1073/pnas.0400628101
M3 - Article
C2 - 15070726
AN - SCOPUS:1842428821
VL - 101
SP - 4379
EP - 4383
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 13
ER -