title: Long-wavelength fluorescent reporters for monitoring protein kinase activity.
abstract: In vivo optical imaging must contend with the limitations imposed by the optical window of tissue (600-1000 nm). 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 600 nm. 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.
authors.affiliation: Department of Chemistry, Division of Chemical Biology and Medicinal Chemistry, and the Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599 (USA).
authors.forename: Nathan P
authors.initials: NP
authors.lastname: Oien
authors.forename: Luong T
authors.initials: LT
authors.lastname: Nguyen
authors.forename: Finith E
authors.initials: FE
authors.lastname: Jernigan
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Priestman
authors.forename: David S
authors.initials: DS
authors.lastname: Lawrence
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry*; D015398:Signal Transduction
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D011494:Protein Kinases
keywords: biosensors; enzymes; fluorescent probes; peptides; signal transduction