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PMID: 18462674 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

X-ray scattering study of activated Arp2/3 complex with bound actin-WCA.

Structure (London, England : 1993) ·Vol. 16 ·No. 5 ·2008-05-00 ·Pages 695-704

Boczkowska M, Rebowski G, Petoukhov MV, Hayes DB, Svergun DI, Dominguez R

Abstract

Previous structures of Arp2/3 complex, determined in the absence of a nucleation-promoting factor and actin, reveal its inactive conformation. The study of the activated structure has been hampered by uncontrollable polymerization. We have engineered a stable activated complex consisting of Arp2/3 complex, the WCA activator region of N-WASP, and one actin monomer, and studied its structure in solution by small angle X-ray scattering (SAXS). The scattering data support a model in which the first actin subunit binds at the barbed end of Arp2, and disqualify an alternative model that places the first actin subunit at the barbed end of Arp3. This location of the first actin and bound W motif constrains the binding site of the C motif to subunits Arp2 and ARPC1, from where the A motif can reach subunits Arp3 and ARPC3. The results support a model of activation that is consistent with most of the biochemical observations.

MeSH Terms
Actin-Related Protein 2-3 Complex/chemistry,isolation & purification,metabolism Actins/chemistry,metabolism Amino Acid Motifs Amino Acid Sequence Animals Brain Chemistry Cattle Mice Models, Molecular Molecular Sequence Data Scattering, Small Angle Sequence Homology, Amino Acid Wiskott-Aldrich Syndrome Protein/chemistry,metabolism X-Ray Diffraction
Chemicals
Actin-Related Protein 2-3 Complex Actins Wiskott-Aldrich Syndrome Protein
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Boczkowska Malgorzata
Department of Physiology, University of Pennsylvania School of Medicine, 3700 Hamilton Walk, A507 Richards Building, Philadelphia, PA 19104-6085, USA.
Rebowski Grzegorz
Petoukhov Maxim V
Hayes David B
Svergun Dmitri I
Dominguez Roberto
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Article Info
Journal
Structure (London, England : 1993)
Abbr.
Structure
ISSN
0969-2126
DOI
10.1016/j.str.2008.02.013
Published
2008-05-00
Pages
695-704
Language
English
Region
United States
NLM ID
101087697
PMCID
PMC2849165
Subset
IM
Grants
NIGMS NIH HHS · R01 GM073791 · United States
NIGMS NIH HHS · R01 GM073791-04 · United States
NIGMS NIH HHS · R01 GM073791-05 · United States
NIGMS NIH HHS · GM073791 · United States
External Links
PubMed source DOI: 10.1016/j.str.2008.02.013 PMC Full Text View journal details
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