BioActive™ Kinase Protein Production

Seattle Genova’s BioActiveTM platform is aiming to offer our customers with excellent protein production services that keep the bioactivities of your proteins just as what they were in nature. To achieve this, we can offer both nature protein purification services and recombinant protein production services. We have developed very specialized protocols to make sure the produced proteins will maintain their biological activities. And our well-developed bioassays can ensure comprehensive activity validations before we release the proteins. Working with a variety of protein expression systems, we are able to rapidly generate bespoke protein reagents to meet customer needs. Backed by years of expertise, we have successfully produced of a variety of proteins for many biological studies.

A protein kinase is a kinase which selectively improves other proteins by covalently adding phosphates to them (phosphorylation) as opposed to kinases which alter lipids, carbohydrates, or other molecules. Phosphorylation usually results in a functional modification of the target protein (substrate) by altering enzyme activity, cellular location, or association with other proteins. The human genome includes about 500 protein kinase genes and they comprise about 2% of all human genes. 

Kinases are significant to drug targets that control cell growth, proliferation, differentiation and metabolism. Drug discovery efforts against kinases remain far from finished. Mutations of various kinases have now been determined to drive several diseases and have also been observed as resistance mechanisms to early 

kinase targeting drug molecules. Thus, for several therapeutically useful kinases with well-established biology, there have already been many “generations” of inhibitors noted to tackle these resistance mechanisms once observed. Recent years have also seen an explosion of interest in kinase drug discovery within academic organizations.

Different kinase Assays 

➢In vitro kinase assays

The choice of which approach you should use depends on the problem you wish to answer, the category of sample to which you have access, and the detection capability you want.

➢Kinase binding assays

This assay can be utilized to study both active and inactive kinases and to evaluate the role of phosphorylation in the interaction with the tracer.

➢Kinase activity assays on ATP-ADP conversion

These categories of assays can be applied to any kinase or an enzyme that converts ATP to ADP as a part of their aactivity.

➢Kinase activity assays based on substrate conversion

An instance is the pyruvate kinase activity assay. This coupled enzyme assay includes two different enzymatic reactions. The early one is the transfer of a phosphate group from phosphoenolpyruvate to ADP by pyruvate kinase, resulting in pyruvate and ATP. The second enzymatic reaction then utilizes pyruvate and a fluorescent peroxidase substrate to develop a fluorescence signal.

The substrate phosphorylation assay

A kinase reaction step is accomplished by firing, and the presence/quantity of phosphorylation is then defined with a phospho-specific antibody.

Workflow

Features of Our Services 

Deliverables

√SDS-PAGE Data

√WB data

√The bioactivity Long-term data 

√The purified proteins are also shipped to our customers for them to test internally

References

1.Manning G, Whyte DB, Martinez R, Hunter T, Sudarsanam S (2002). "The protein kinase complement of the human genome". Science. 298 (5600): 1912–1934.

2.Besant PG, Tan E, Attwood PV (March 2003). "Mammalian protein histidine kinases". Int. J. Biochem. Cell Biol. 35 (3): 297–309.

3.Roskoski, R., Jr. Assays of protein kinase. Methods Enzymol. 99, 3–6 (1983).