Cell-permeable probe for identification and imaging of sialidases.

Charng-Sheng Tsai, Hsin-Yung Yen, Meng-I Lin, Tsung-I Tsai, Shi-Yun Wang, Wen-I Huang, Tsui-Ling Hsu, Yih-Shyun E Cheng, Jim-Min Fang, Chi-Huey Wong

Proceedings of the National Academy of Sciences of the United States of America 2013 Feb 12

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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.

Citation

Charng-Sheng Tsai, Hsin-Yung Yen, Meng-I Lin, Tsung-I Tsai, Shi-Yun Wang, Wen-I Huang, Tsui-Ling Hsu, Yih-Shyun E Cheng, Jim-Min Fang, Chi-Huey Wong. Cell-permeable probe for identification and imaging of sialidases. Proceedings of the National Academy of Sciences of the United States of America. 2013 Feb 12;110(7):2466-71