First Synthetic Probe for the Detection and Quantification of a Protein with a Potential α,(2→8)Sialyltransferase Activity

Autor:	Arduino Oratore, and Anna Rita Lizzi, Gabriele D'Andrea
Rok vydání:	2004
Předmět:	Pharmacology biology Sialyltransferase Chemistry Organic Chemistry Biomedical Engineering Proteins Pharmaceutical Science A protein Bioengineering Sialyltransferases Sialyltransferase activity Biochemistry Polysaccharides Cell Line Tumor Molecular Probes biology.protein Humans Biotechnology
Zdroj:	Bioconjugate Chemistry. 15:1084-1087
ISSN:	1520-4812 1043-1802
DOI:	10.1021/bc049970s
Popis:	There is considerable interest in monitoring alpha,(2--8)sialyltransferase (ST8) levels; however, there are few specific and sensitive methods to directly detect and quantitate the protein. This paper reports the development of a synthetic probe composed of oxidized colominic acid coupled to biotinyl-L-lysine hydrazide to detect and quantify ST8 with putative "initiase" activity and its use in three solid-phase applications. The detection limit observed for ST8 purified from K562 cells was approximately 2 pg by dot-blot analysis. In Western blots the probe bound and specifically recognized a protein band corresponding to ST8. In ELISA a linear dose response was obtained for pure protein in the range of 50-200 pg. Analysis of 3'-azido-3'deoxythymidine-treated cells by all three methods showed a reduction in ST8 compared to control cells; treated cells had 73% of control levels by ELISA. This probe will be useful for studies on the expression ST8 and its role in glycoconjugate biosynthesis.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::68baf44aa095207b3c2c15622334b5f8 https://doi.org/10.1021/bc049970s Zobrazit plný text záznamu