Peptide-based kinase reporters: Design and caveats

Authors

Alen Piljic, Carsten Schultz

Abstract

Phosphorylation and dephosphorylation by kinases and phosphatases, respectively, are considered the most frequent posttranslational protein modifications. The human genome hosts about 540 kinases and a slightly smaller number of phosphatases. In order to monitor their activity, fluorescent sensors are highly desirable tools. In this chapter, we describe our attempts to prepare small peptide-based fluorescent sensors for one of the most common protein kinases. Synthetic pathways and coupling strategies for attaching the fluorophore were developed. The resulting labeled peptides were then used in enzyme-based assays. What initially looked as being the perfect solution for our goal, turned out to be a pure artifact. The results demonstrate some of the difficulties in preparing a successful small molecule-based sensor and the need of proper controls, especially for the biological testing.

Documents for download

• Piljic_BookChapter_final_1.pdf

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