Alkylation of staurosporine to derive a kinase probe for fluorescence applications
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...
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| Format: | Article |
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Wiley-VCH Verlag
2016
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| Online Access: | https://eprints.nottingham.ac.uk/40837/ |
| _version_ | 1848796144001351680 |
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| author | Disney, Alexander J.M. Kellam, Barrie Dekker, Lodewijk V. |
| author_facet | Disney, Alexander J.M. Kellam, Barrie Dekker, Lodewijk V. |
| author_sort | Disney, Alexander J.M. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:43:18Z |
| format | Article |
| id | nottingham-40837 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:43:18Z |
| publishDate | 2016 |
| publisher | Wiley-VCH Verlag |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-408372020-05-04T17:40:48Z https://eprints.nottingham.ac.uk/40837/ Alkylation of staurosporine to derive a kinase probe for fluorescence applications Disney, Alexander J.M. Kellam, Barrie Dekker, Lodewijk V. 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. Wiley-VCH Verlag 2016-03-23 Article PeerReviewed Disney, Alexander J.M., Kellam, Barrie and Dekker, Lodewijk V. (2016) Alkylation of staurosporine to derive a kinase probe for fluorescence applications. ChemMedChem, 11 . pp. 972-979. ISSN 1860-7187 http://onlinelibrary.wiley.com/doi/10.1002/cmdc.201500589/abstract doi:10.1002/cmdc.201500589 doi:10.1002/cmdc.201500589 |
| spellingShingle | Disney, Alexander J.M. Kellam, Barrie Dekker, Lodewijk V. Alkylation of staurosporine to derive a kinase probe for fluorescence applications |
| title | Alkylation of staurosporine to derive a kinase probe for fluorescence applications |
| title_full | Alkylation of staurosporine to derive a kinase probe for fluorescence applications |
| title_fullStr | Alkylation of staurosporine to derive a kinase probe for fluorescence applications |
| title_full_unstemmed | Alkylation of staurosporine to derive a kinase probe for fluorescence applications |
| title_short | Alkylation of staurosporine to derive a kinase probe for fluorescence applications |
| title_sort | alkylation of staurosporine to derive a kinase probe for fluorescence applications |
| url | https://eprints.nottingham.ac.uk/40837/ https://eprints.nottingham.ac.uk/40837/ https://eprints.nottingham.ac.uk/40837/ |