Alkylation of staurosporine to derive a kinase probe for fluorescence applications

Disney, Alexander J.M.; Kellam, Barrie; Dekker, Lodewijk V.

Abstract

The natural product staurosporine is a high-affinity inhibitor of nearly all mammalian protein kinases.The labelling of staurosporine has proven effective as a means of generating protein kinase research tools. Most tools have been generated by acylation of the 4’-methylamine of the sugar moiety of staurosporine. Herein we describe the alkylation of this group as a first step to generate a fluorescently labelled staurosporine. Following alkylation, a polyethylene glycol linker was installed, allowing subsequent attachment of fluorescein. We report that this fluorescein–staurosporine conjugate binds to cAMP-dependent protein kinase in the nanomolar range. Furthermore, its binding can be antagonised with unmodified staurosporine as well as ATP, indicating it targets the ATP binding site in a similar fashion to native staurosporine. This reagent has potential application as a screening tool for protein kinases of interest.

Citation

Disney, A. J., Kellam, B., & Dekker, L. V. (in press). Alkylation of staurosporine to derive a kinase probe for fluorescence applications. ChemMedChem, 11, https://doi.org/10.1002/cmdc.201500589

• Journal Article Type: Article
• Acceptance Date: Feb 10, 2016
• Online Publication Date: Mar 23, 2016
• Deposit Date: Feb 27, 2017
• Publicly Available Date: Feb 27, 2017
• Journal: ChemMedChem
• Print ISSN: 1860-7179
• Electronic ISSN: 1860-7187
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 11
• DOI: https://doi.org/10.1002/cmdc.201500589
• Public URL: https://nottingham-repository.worktribe.com/output/779385
• Publisher URL: http://onlinelibrary.wiley.com/doi/10.1002/cmdc.201500589/abstract
• Contract Date: Feb 27, 2017