Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 53, Issue 15, Pages 3975-3978Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201309691
Keywords
biosensors; enzymes; fluorescent probes; peptides; signal transduction
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Funding
- National Institutes of Health [RO1 CA79954]
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Invivo optical imaging must contend with the limitations imposed by the optical window of tissue (600-1000nm). Although a wide array of fluorophores are available that are visualized in the red and near-IR region of the spectrum, with the exception of proteases, there are few long wavelength probes for enzymes. This situation poses a particular challenge for studying the intracellular biochemistry of erythrocytes, the high hemoglobin content of which optically obscures subcellular monitoring at wavelengths less than 600nm. To address this, tunable fluorescent reporters for protein kinase activity were developed. The probing wavelength is preprogrammed by using readily available fluorophores, thereby enabling detection within the optical window of tissue, specifically in the far-red and near-IR region. These agents were used to monitor endogenous cAMP-dependent protein kinase activity in erythrocyte lysates and in intact erythrocytes when using a light-activatable reporter.
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