Product Pathways - Kinases
Ack1 Kinase #7426
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Description
Purified recombinant human Ack1 (Gly110-Trp476) kinase, supplied as a GST fusion protein.
Source / Purification
The GST-Kinase fusion protein was produced using a baculovirus expression system with a construct expressing a fragment of human Ack1 (Gly110-Trp476) (GenBank Accession No. NM_005781.4) 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-Ack1 fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. Ack1 kinase activity was measured in a radiometric assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 μM Na-orthovanadate, 1.2 mM DTT, 2.5 μg/50 μl PEG20,000, ATP variable, Substrate: Poly (EY. 4:1) 20 ng/μl and recombinant Ack1: 200 ng/50 μl.
Quality Control
The theoretical molecular weight of the GST-Ack1 fusion protein is 71 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. Ack1 kinase activity was determined using a radiometric assay [Fig.2].
Background
Ack1 and Ack2 (activated cdc42-associated kinase 1 and 2) are non-receptor tyrosine kinases that consist of a tyrosine kinase core, an SH3 domain, a cdc42/Rac-binding (CRIB) domain, a Ralt homology region and a proline-rich region (1,2). Ack1 and 2 are the only two tyrosine kinases known to interact with cdc42. Both Acks are activated by growth factors including EGF and PDGF, as well as by activated integrins through cell adhesion, and may serve to link receptor tyrosine kinase or G protein-coupled receptor signaling with cdc42. Acks may regulate cell growth, morphology and motility (3,4). Recent findings indicate that Ack1 may play a role in prostate tumorigenesis, making it a potential drug target for this type of cancer (5).
- Galisteo, M.L. et al. (2006) Proc. Natl. Acad. Sci. USA 103, 9796-9801.
- Yokoyama, N. and Miller, W.T. (2003) J. Biol. Chem. 278, 47713-47723.
- Yang, W. and Cerione, R.A. (1997) J. Biol. Chem. 272, 24819-24824.
- Yang, W. et al. (2001) J. Biol. Chem. 276, 43987-43993.
- Mahajan, N.P. et al. (2005) Cancer Res. 65, 10514-10523.
Application References
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Companion Products
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.
