Chemistry

Bifunctional Molecular Probes for Activity-Based Visualization of Quinone-Dependent Amine Oxidases

Document Type

Article

Abstract

The design, synthesis, and evaluation of two bifunctional molecular probes that can be used to visualize quinone-dependent amine oxidase enzymes in an activity-dependent manner are described. These probes use alkylhydrazines to irreversibly bind the target enzymes, which can then be visualized with either Western blotting or in-gel fluorescence. The results show that the Western blotting readout, which utilizes commercially available anti-nitrophenyl antibodies to detect a simple dinitrophenyl antigen, provides a stronger readout than the fluorescein-based fluorescence readout. This visualization strategy can be used to measure the potency of enzyme inhibitors by selectively visualizing the active enzyme that remains after treatment with an inhibitor. Looking forward, this probe molecule and visualization strategy will enable activity-based protein-profiling experiments, such as determining inhibitor selectivity values within full proteome mixtures, for this family of amine oxidase enzymes.

Publication Title

Chemistry - A European Journal

Publication Date

12-3-2018

Volume

24

Issue

67

First Page

17681

Last Page

17685

ISSN

0947-6539

DOI

10.1002/chem.201804247

Keywords

arenes, immunoassays, inhibitors, protein modifications, proteomics

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