Vinculin, phosphorylated (Tyr1065)

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 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. Phosphorylation of vinculin at pY1065 is c-Src dependent. This has been observed in vitro and in activated platelets. Phosphorylation of vinculin on tyrosine residue 1065 has been shown to affect head-tail interactions and play a role in cell spreading; however, it does not affect the interaction between vinculin and actin or cellular localization.\n\nApplications: \nSuitable for use in Western Blot following Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000 \nOptimal dilutions to be determined by the researcher.\n\nPositive Control:\nCOS cells co-transfected with activated Src and His-tagged chicken vinculin cDNA were treated with vanadate for 24 hr.\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

V2122-74

Size

10 Blots

Applications

IP, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, 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. Zhang, Z., et al. (2004) The phosphorylation of vinculin on tyrosine residues 100 and 1065, mediated by SRC kinases, affects cell spreading. Mol. Biol. Cell. 15(9):4234-4247. 2. Subauste, M.C., et al. (2004) Vinculin modulation of paxillin-FAK interactions regulates ERK to control survival and motility. J. Cell. Biol. 165(3):371-381. 3. 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. 4. 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. 5. Ito, S., et al. (1983) Vinculin phosphorylation by the src kinase. Interaction of vinculin with phospholipid vesicles. J. Biol. Chem. 258(23):14626-14631.

Additional Info

Recognizes chicken and human Vinculin when phosphorylated at Tyr1065. Species sequence homology: Mouse vinculin (100%).