University of Southern California

Sayantan Mitra

Amy M Barrios

Nunzio Bottini

Stephanie Stanford

The design, synthesis, and utilization of a novel coumarin-based amino acid and its fluorinated derivatives for use in enzyme activity and substrate specificity assays are described. The synthesis is both facile and high-yielding and allows for the production of a highly fluorescent phosphorylated amino acid that can be incorporated into peptides using standard SPPS techniques. The resultant substrates are efficiently hydrolyzed by PTPs and exhibit a large increase in fluorescence upon hydrolysis. Fluorinated CAP derivatives have utility as fluorogenic enzyme substrates. CAP and its derivatives can be incorporated into combinatorial peptide libraries for assaying enzyme substrate specificities.

Coumarin-based amino acids for use in enzyme activity and substrate specificity assays

The invention relates in general to tyrosine dephosphorylation. More specifically, the invention relates to methods and compositions for monitoring intracellular tyrosine dephosphorylation at the single cell level. The invention further relates to techniques that can be used as aid in the development of novel therapeutics, and monitor regulation of intracellular tyrosine phosphatase activity at the single cell level.

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Los Angeles, CA, United States of America

Sayantan Mitra