Design and synthesis of an ER-specific fluorescent probe based on carboxylesterase activity with quinone methide cleavage process

Bioorganic and Medicinal Chemistry Letters

Volumn 21, Issue 11, 2011, Pages 3206-3209

  Hakamata, Wataru ^a   MacHida, Aki ^a   Oku, Tadatake ^a   Nishio, Toshiyuki ^a  

^a NIHON UNIVERSITY   (Japan)

Author keywords

Carboxylesterase; Fluorescent probe; Quinone methide cleavage; Resorufin

Indexed keywords

CARBOXYLESTERASE; FLUORESCENT DYE; QUINONE DERIVATIVE;

ANIMAL CELL; ARTICLE; CELL DIVISION; CELL LEVEL; CONTROLLED STUDY; DRUG DESIGN; DRUG SYNTHESIS; ENDOPLASMIC RETICULUM; FLUORESCENCE; HUMAN; HUMAN CELL; HYDROLYSIS; NONHUMAN;

ANIMALS; CARBOXYLESTERASE; CERCOPITHECUS AETHIOPS; COS CELLS; DRUG DESIGN; ENDOPLASMIC RETICULUM; FLUORESCENT DYES; INDOLEQUINONES; MOLECULAR STRUCTURE;

EID: 79956097639     PISSN: 0960894X     EISSN: 14643405     Source Type: Journal    
DOI: 10.1016/j.bmcl.2011.04.066     Document Type: Article

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9. Photochemical characterization. The solutions of resorufin and probe 1 (10 μM) in PBS(-) containing 0.1% DMSO, respectively. The fluorescence emission spectra of these solutions were recorded with an excitation wavelength of 571 nm.
10. Enzyme assays. Probe 1 was prepared as a 1 mM solution in DMSO. Esterase (from porcine liver, purchased from SIGMA) was prepared as a 16.5 μg/mL solution in 200 mM Na phosphate buffer (pH 8.0). The assay were conducted by adding esterase solution (85 μL) to 1 (5 μL) and 200 mM Na phosphate buffer, pH 8.0 (100 μL) followed by incubation at 37 °C. The assays were followed by the monitoring fluorescence intensity change of resorufin. One unit of enzyme activity was defined as the amount of enzyme required to liberate 1 μmol of resorufin/min.
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