A new technology for functional solubilization of kinases

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Department of Health and Human Services
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Phase I
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6201 La Pas Trail, SUITE 160, INDIANAPOLIS, IN, 46268
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DESCRIPTION (provided by applicant): Protein kinases are highly important enzymes that function primarily as components of signaling pathways, regulating cellular responses such as cell growth, development, differentiation, membrane transport, and cell dea th. Abnormalities in these signaling pathways can lead to various pathological conditions including many forms of cancer. For these reasons, protein kinases are important targets for both basic research and drug development. However, structural and funct ional analyses of cancer-associated kinases are often challenging or even infeasible due to poor protein solubility. Although several approaches have been developed for improving recombinant protein solubility, many kinases remain recalcitrant to these so lubilization techniques. Therefore, there is a high demand in novel technologies that increase the chances for the soluble expression of recombinant cancer-associated kinases that were not soluble with other technologies. This project aims to imple ment our knowledge of the protein intrinsic disorder phenomenon for the development of a novel solubilization approach based on the fusion of target cancer-related kinases to highly flexible, non-aggregating polypeptide chains, known as entropic bristle do mains (EBDs). Phase I of this project is dedicated to demonstrating that a known insoluble kinases can be functionally solubilized by the EBD fusion. This goal will be achieved via the following specific aims: (1) demonstrate that a known insoluble kinas e can be solubilized by the EBD fusion; and (2) demonstrate that the EBD fusion does not affect the kinase functionality. The final envisioned product of this proposal is a set of EBD vectors, brand named AquoKinTM vectors that can be used to incre ase the functional solubility of recombinant kinases. AquoKinTM will benefit any and all researchers needing to increase kinase solubility, whether their research involves specifically cancer-related target or other disease-related kinases. Since kinases are often being studied by anti-cancer researchers, AquoKinTM will expand the number of kinases targets that can be studied in the fight against cancer.

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