Product Pathways - Kinases
Syk Kinase #7368
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Description
Purified recombinant full-length human Syk (Met1-Asn635) 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 full-length human Syk (Met1-Asn635) (GenBank Accession No. NM_003177) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using glutathione-agarose.
Gel Staining
Figure 1. The purity of the GST-Syk fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. Syk 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, 4 mM MgCl2, 2.5 mM MnCl2, 0.05 mM DTT, 50 μM ATP, Substrate: Poly EY, 400 ng/μL, and recombinant Syk: variable.
Quality Control
The theoretical molecular weight of the GST-Syk fusion protein is 100 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. Syk kinase activity was determined using a radiometric assay [Fig.2].
Background
Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation and phagocytosis (4). There is also evidence of a role for Syk in nonimmune cells, and Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation of Tyr323 provides a direct binding site to the TKB domain of Cbl (6,7). Tyrosine 352 of Syk is involved in the association of PLC-γ1 (8). Tyrosines 525 and 526 are located in the activation loop of the Syk kinase domain, and phosphorylation of Tyr525/526 of human Syk (equivalent to the Tyr519/520 of mouse Syk) is essential for Syk function (9).
- Cheng, A.M. and Chan, A.C. (1997) Curr. Opin. Immunol. 9, 528-533.
- Kurosaki, T. et al. (1997) Curr. Opin. Immunol. 9, 309-318.
- Chu, D.H. et al. (1998) Immunol. Rev. 165, 167-180.
- Turner, M. et al. (2000) Immunol. Today 21, 148-154.
- Coopman, P.J. et al. (2000) Nature 406, 742-747.
- Decker, M. et al. (1998) J. Biol. Chem. 273, 8867-8874.
- Rao, N. et al. (2001) EMBO J. 20, 7085-7095.
- Law, C.L. et al. (1996) Mol. Cell. Biol. 16, 1305-1315.
- Zhang, J. et al. (2000) J. Biol. Chem. 275, 35442-35447.
Application References
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Companion Products
- 9805 HTScan® Tyrosine Kinase Buffer (4X)
- 7404 RSK2 Kinase
- 7450 Tyrosine Kinase Substrate Screening Kit
- 7779 HTScan® Syk Kinase Assay Kit
- 1310 Gastrin Precursor (Tyr87) Biotinylated Peptide
- 9411 Phospho-Tyrosine Mouse mAb (P-Tyr-100)
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.
