NMR analyses of the activation of the Arp2/3 complex by neuronal Wiskott-Aldrich syndrome protein

Mara Kreishman-Deitrick, Erin D. Goley, Lyle Burdine, Carilee Denison, Coumaran Egile, Rong Li, Nagarajan Murali, Thomas J. Kodadek, Matthew D. Welch, Michael K. Rosen

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39 Scopus citations

Abstract

The VCA domain of the neuronal Wiskott-Aldrich syndrome protein (N-WASP) is a potent activator of the Arp2/3 complex, a 240 kDa heteroheptameric actin-nucleating assembly. We used site-directed spin labeling of N-WASP peptides in conjunction with methyl-TROSY spectra of the intact, selectively labeled Arp2/3 complex to identify regions of the VCA that are proximal to the ARPC3 subunit of the assembly. We also cross-linked CA peptides to the Arp3, Arp2, ARPC1, and ARPC3 subunits. The combined data suggest that the extreme C-terminus of the A region and the C-terminus of the C region of N-WASP are proximal to ARPC3. These results have implications for the mechanism of Arp2/3 complex activation by VCA peptides. This study also demonstrates the utility of NMR spectroscopy for studying ligand binding events in large, asymmetric, macromolecular assemblies.

Original languageEnglish (US)
Pages (from-to)15247-15256
Number of pages10
JournalBiochemistry
Volume44
Issue number46
DOIs
StatePublished - Nov 22 2005

ASJC Scopus subject areas

  • Biochemistry

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    Kreishman-Deitrick, M., Goley, E. D., Burdine, L., Denison, C., Egile, C., Li, R., Murali, N., Kodadek, T. J., Welch, M. D., & Rosen, M. K. (2005). NMR analyses of the activation of the Arp2/3 complex by neuronal Wiskott-Aldrich syndrome protein. Biochemistry, 44(46), 15247-15256. https://doi.org/10.1021/bi051065n