Product Pathways - Kinases
Fes Kinase #7643
Cell Signaling Technology offers a full line of protein kinases, substrates, antibody detection reagents and HTScan® kits. Browse our "Reagents for High-Throughput Screening" product listing or contact us at drugdiscovery@cellsignal.com.
Description
Purified recombinant full-length human Fes kinase, supplied as a GST fusion protein.
Source / Purification
The GST-Kinase fusion protein was produced using a baculovirus expression system using sf9 cells and a recombinant virus encoding full length human Fes (Met1-Arg822) (GenBank Accession No. NM_002005) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using GSH-agarose.
Gel Staining
Figure 1. The purity of the GST-Fes fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. Fes kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 5 mM MgCl2, 0.05 mM DTT, 50 μM ATP, Substrate: Poly (Glu:Tyr, 4:1) 400 ng/μL, and recombinant Fes: variable.
Quality Control
The theoretical molecular weight of the GST-Fes fusion protein is 125 kDa. The purity of the kinase was assessed using SDS-PAGE followed by Coomassie stain [Fig.1]. Fes kinase activity was determined using a radiometric assay [Fig.2].
Background
Fes/Fps and Fer are the only two members of a unique family of cytoplasmic protein tyrosine kinases (1,2). Fps and Fer contain a central Src homology-2 (SH2) domain and a carboxy-terminal tyrosine kinase catalytic domain. They are structurally distinguished from other members of cytoplasmic protein tyrosine kinase subfamilies by the presence of amino-terminal Fer/CIP4 homology and coiled-coil domains (3). Fes/Fps was originally identified as an oncogene from avian (Fps) and feline (Fes) retroviruses. Human c-Fes has been implicated in myeloid, vascular endothelial and neuronal cell differentiation. Mutations may activate the Fps kinase and thereby contribute to cancer (4). However, recent data strongly suggests that the c-Fes protein-tyrosine kinase is a tumor suppressor rather than a dominant oncogene in colorectal cancer (5).
- Smithgall, T.E. et al. (1998) Crit. Rev. Oncog. 9, 43-62.
- Greer, P. (2002) Nat. Rev. Mol. Cell Biol. 3, 278-289.
- Sangrar, W. et al. (2005) Cancer Res. 65, 3518-3522.
- Ley, T.J. et al. (2003) Proc Natl Acad Sci U S A 100, 14275-14280.
- Delfino, F.J. et al. (2006) J. Biol. Chem. 281, 8829-8835.
Application References
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Companion Products
- 9805 HTScan® Tyrosine Kinase Buffer (4X)
- 9804 ATP (10 mM)
- 7450 Tyrosine Kinase Substrate Screening Kit
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
