Small G-proteins
RhoGDI protein, GST-tagged

Cat. # GDI01

Product Uses Include

  • In vitro binding and inhibition of prenylated Rho proteins
  • RhoGDI protein interaction studies

Material
Rho gunanine nucleotide dissociation inhibitor-1 (RhoGDI) binds with high affinity to Cdc42, Rac and Rho family members that have been post-translationally modified at their carboxy-terminal with a prenyl group and with a much lower affinity to the corresponding non-prenylated proteins. RhoGDI is thought to solubilize the prenylated proteins and regulate their association with cellular membranes. Recombinant RhoGDI is supplied as a lyophilized powder. Reconstitution of the protein in water to 1 mg/ml leaves the protein in the following buffer: 2 mM Tris pH 7.6, 0.5 mM MgCl2, 0.5% sucrose and 0.1% dextran. Protein concentration is determined by the Precision Red Advanced Protein Assay Reagent (Cat. # ADV02).

Recombinant RhoGDI has been expressed as a GST fusion protein in a bacterial system. The recombinant protein is 55 kDa in size, consisting of the RhoGDI protein (27 kDa) and an amino-terminal 28 kDa GST tag.

Purity
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are 90% pure.

Figure 1: GST-RhoGDI protein purity determination. A 20 µg sample of GDI01 was separated by electrophoresis in a 12% SDS-PAGE system. The protein was stained with Coomassie Blue.

Biological Activity
Biological activity of GST-RhoGDI protein was determined by the ability of the protein to inhibit GDP exchange on geranylgeranylated Cdc42

Examples of publications where this product was used:
Guo, Z., Liu, L., Cafiso, D. and Castle, D. (2002). Perturbation of a very late step of regulated exocytosis by a secretory carrier membrane protein (SCAMP2)-derived peptide. J. Biol. Chem. 277, 35357-35363.

Mehta, D., Rahman, A. and Malik, A. B. (2001). Protein kinase C-α signals rho-guanine nucleotide dissociation inhibitor phosphorylation and rho activation and regulates the endothelial cell barrier function. J. Biol. Chem. 276, 22614-22620.

Yamashita, T. and Tohyama, M. (2003). The p75 receptor acts as a displacement factor that releases Rho from Rho-GDI. Nat. Neurosci. 6, 461-467.
Product description Cat. # Amount Price & Order
RhoGDI protein, GST-tagged GDI01-A 1 x 25 µg
GDI01-B 4 x 25 µg