Vinculin, phosphorylated (Tyr822)

Vinculin is an ubiquitously expressed cytoskeletal protein (~130kD) involved in cell adhesion and cell migration. The vinculin protein consists of a globular head domain connected to an elongated tail region by a proline-rich domain. The head region contains binding sites for two cytoskeletal proteins, a-actinin and talin, as well as a binding site for the tail region of vinculin itself. The linker region possesses binding sites for two adaptor proteins, vinexin and ponsin, that may connect growth factor receptors with integrin signaling pathways, and a binding site for an actin-bundling protein, VASP. The tail region contains binding sites for actin, the cytoskeletal protein, paxillin, and PI(4,5)P2. In the inactive state the head region of vinculin is bound to the tail region, resulting in inaccessibility of the other protein binding sites. Binding of PI(4,5)P2 releases the head-tail interaction allowing binding of other proteins to vinculin. These regulatory events play an important role in the formation, maintenance, and breakdown of focal adhesions that occur during cell adhesion and migration. In addition to these protein binding sites, the head and tail regions of vinculin have multiple potential phosphorylation sites. Data indicate the possible involvement of phosphorylation of tyrosine 822 of vinculin with the integrity/strength of integrinmediated focal adhesions.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000 \nOptimal dilutions to be determined by the researcher.\n\nPositive Control:\nChick Embryo Fibroblasts (CEFs) transfected with activated Src\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

V2122-72

Size

10 Blots

Applications

WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat, Ch/Bird

Form

Supplied as a liquid in PBS (without Mg2+ and Ca2+), 1mg/ml BSA (IgG, protease free), pH 7.2, 0.05% sodium azide, 50% glycerol.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

References

1. Opaza Saez, A.M., et al. (2003) Tension development during contractile stimulation of smooth muscle requires recruitment of paxillin and vinculin to the membrane. Am. J. Physiol. Cell Physiol. (ePub ahead of print; October 22, 2003). 2. von Wichert, G., et al. (2003) Force-dependent integrin-cytoskeleton linkage formation requires downregulation of focal complex dynamics by Shp2. EMBO J. 22(19):5023-5035. 3. Zaidel-Bar, R., et al. (2003) Early molecular events in the assembly of matrix adhesions at the leading edge of migrating cells. J. Cell Sci. 116(Pt 22):4605-4613. 4. Ziegler, W.H., et al. (2002) A lipid-regulated docking site on vinculin for protein kinase C. J. Biol. Chem. 277(9):7396-7404. 5. Zamir, E. and B. Geiger (2001) Molecular complexity and dynamics of cell-matrix adhesions. J. Cell Sci. 114(Pt 20):3583-3590. 6. Sechi, A.S. and J. Wehland (2000) The actin cytoskeleton and plasma membrane connection: PtdIns(4,5)P2 influences cytoskeletal protein activity at the plasma membrane. J. Cell Sci. 113(Pt 21):3685-3695. 7. Goldmann, W.H., et al. (1998) Differences in elasticity of vinculin-deficient F9 cells measured by magnetometry and atomic force spectroscopy. Exp. Cell Res. 239(2):235-242.

Additional Info

Recognizes chicken vinculin when phosphorylated at Tyr822. Species sequence homology: human, mouse, and rat vinculin (100%).