Cell-permeable probe for identification and imaging of sialidases
Journal
National Academy of Sciences of the United States of America
Journal Volume
110
Journal Issue
7
Pages
2466-2471
Date Issued
2013
Author(s)
Tsai, C.-S.
Yen, H.-Y.
Lin, M.-I.
Tsai, T.-I.
Wang, S.-Y.
Huang, W.-I.
Hsu, T.-L.
Cheng, Y.-S.E.
Wong, C.-H.
Abstract
Alkyne-hinged 3-fluorosialyl fluoride (DFSA) containing an alkyne group was shown to be a mechanism-based target-specific irreversible inhibitor of sialidases. The ester-protected analog DFSA (PDFSA) is a membrane-permeable precursor of DFSA designed to be used in living cells, and it was shown to form covalent adducts with virus, bacteria, and human sialidases. The fluorosialyl-enzyme adduct can be ligated with an azide-annexed biotin via click reaction and detected by the streptavidin-specific reporting signals. Liquid chromatography-mass spectrometry/mass spectrometry analysis on the tryptic peptide fragments indicates that the 3-fluorosialyl moiety modifies tyrosine residues of the sialidases. DFSA was used to demonstrate influenza infection and the diagnosis of the viral susceptibility to the anti-influenza drug oseltamivir acid, whereas PDFSA was used for in situ imaging of the changes of sialidase activity in live cells.
Type
journal article