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The EVH2 Domain of the Vasodilator-stimulated Phosphoprotein Mediates Tetramerization, F-actin Binding, and Actin Bundle Formation
Open accessAbstract
Vasodilator-stimulated phosphoprotein (VASP) is a member of the Ena/VASP family of proteins that are implicated in regulation of the actin cytoskeleton. All family members share a tripartite structural organization, comprising an N-terminal Ena/VASP homology (EVH) 1 domain, a more divergent proline-rich central part, and a common C-terminal EVH2 region of about 160-190 amino acids. Using chemical cross-linking, sucrose gradient sedimentation, and gel filtration analyses of different truncated VASP constructs, we demonstrate that the VASP EVH2 region is both necessary and sufficient for tetramerization. Moreover, co-sedimentation and fluorescent phalloidin staining showed that the EVH2 region binds and bundles F-actin in vitro and localizes to stress fibers in transfected cells. Analysis of the functional contribution of highly conserved blocks within this region indicated that residues 259-276 of human VASP are essential for the interaction with F-actin, whereas residues 343-380 are required for tetramerization, probably via coiled-coil formation. Interactions with F-actin are enhanced by VASP tetramerization. The results demonstrate that the C-terminal EVH2 segment is not only conserved in sequence but also forms a distinct functional entity. The data suggest that the EVH2 segment represents a novel oligomerization and F-actin binding domain.
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Stoscheck, C. M. (1990) · Methods in Enzymology · doi:10.1016/0076-6879(90)82008-p · Cited by 1,438
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Metadata
- Authors
- Bachmann, C., Fischer, L., Walter, U., & Reinhard, M.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1999-08
- Volume
- 274
- Issue
- 33
- Pages
- 23549-23557
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2077489546
- Cited by
- 293
- References
- 2
- FWCI
- 5.90
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Bachmann, C., Fischer, L., Walter, U., & Reinhard, M. (1999). The EVH2 Domain of the Vasodilator-stimulated Phosphoprotein Mediates Tetramerization, F-actin Binding, and Actin Bundle Formation. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.274.33.23549
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