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Accepted (AM)Open Access (License Unspecified), Open
Book chapter
Biochemical Reconstitution of the WAVE Regulatory Complex
Methods in Enzymology, pp 55-72
01 Jan 2014
PMID: 24630101
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
The WAVE regulatory complex (WRQ is a 400-kDa heteropentameric protein assembly that plays a central role in controlling actin cytoskeletal dynamics in many cellular processes. The WRC acts by integrating diverse cellular cues and stimulating the actin nucleating activity of the Arp2/3 complex at membranes. Biochemical and biophysical studies of the underlying mechanisms of these processes require large amounts of purified WRC. Recent success in recombinant expression, reconstitution, purification, and crystallization of the WRC has greatly advanced our understanding of the inhibition, activation, and membrane recruitment mechanisms of this complex. But many important questions remain to be answered. Here, we summarize and update the methods developed in our laboratory, which allow reliable and flexible production of tens of milligrams of recombinant WRC of crystallographic quality, sufficient for many biochemical and structural studies.
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Details
- Title
- Biochemical Reconstitution of the WAVE Regulatory Complex
- Creators
- Baoyu Chen - The University of Texas Southwestern Medical CenterShae B. Padrick - The University of Texas Southwestern Medical CenterLisa Henry - The University of Texas Southwestern Medical CenterMichael K. Rosen - The University of Texas Southwestern Medical Center
- Contributors
- R D Vale (Editor)
- Publication Details
- Methods in Enzymology, pp 55-72
- Series
- Methods in Enzymology
- Publisher
- Elsevier; SAN DIEGO
- Number of pages
- 18
- Grant note
- R01-GM056322; R01 GM056322 / NIGMS NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) Howard Hughes Medical Institute R01GM056322 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS)
- Resource Type
- Book chapter
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000334128900005
- Scopus ID
- 2-s2.0-84896342786
- Other Identifier
- 991020836334004721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology