Premium
A New Fluorogenic Probe for the Detection of endo ‐β‐ N ‐Acetylglucosaminidase
Author(s) -
Ishii Nozomi,
Sunaga Chie,
Sano Kanae,
Huang Chengcheng,
Iino Kenta,
Matsuzaki Yuji,
Suzuki Tadashi,
Matsuo Ichiro
Publication year - 2018
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201700662
Subject(s) - chemistry , fluorescence , hydrolysis , derivative (finance) , combinatorial chemistry , high throughput screening , quenching (fluorescence) , chromatography , stereochemistry , biochemistry , physics , quantum mechanics , financial economics , economics
We developed a fluorescence‐quenching‐based assay system to determine the hydrolysis activity of endo ‐β‐ N ‐acetylglucosaminidases (ENGases). The pentasaccharide derivative 1 was labeled with an N ‐methylanthraniloyl group as a reporter dye at the non‐reducing end and with a 2,4‐dinitrophenyl group as a quencher molecule at the reducing end. This derivative is hydrolyzed by ENGase, resulting in an increase in fluorescence intensity. Thus, the fluorescence signal is directly proportional to the amount of the tetrasaccharide derivative, hence allowing ENGase activity to be evaluated easily and quantitatively. Using this system, we succeeded in measuring the hydrolysis activities of ENGases and thus the inhibitory activities of known inhibitors. We confirmed that this assay system is suitable for high‐throughput screening for potential inhibitors of human ENGase that might serve as therapeutic agents for the treatment of N ‐glycanase 1 ( NGLY1 ) deficiency.