Abstract
Determining glycosyltransferase activities gives a clue for better understanding an underlying mechanism for glycomic alterations of carrier molecules. N-glycan branch formation is concertedly regulated by cooperative and competitive activities of N-acetylglucosaminyltransferases (GnTs). Here, we describe methods for large scale preparation of the oligosaccharide acceptor substrate, fluorescence-labeling of oligosaccharides by pyridylamination, quality control, and reversed phase HPLC-based measurement of GnT activities including GnT-III, IV, V, and IX.
Authors Shinji Takamatsu and Hiroaki Korekane have contributed equally to this work.
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Acknowledgments
This work was mainly supported by Grant-in-ÂAid for Scientific Research (A), No. 20249018, and the Global COE Program of Osaka University funded by the Ministry of Education, Culture, Sports, Science, and Technology of Japan, the Naito foundation, and the National Institute of Biomedical Innovation (NIBIO) in Japan. We also thank Ms. Fumi Ota for supporting in the preparation of this manuscript.
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Takamatsu, S. et al. (2013). N-Acetylglucosaminyltransferase (GnT) Assays Using Fluorescent Oligosaccharide Acceptor Substrates: GnT-III, IV, V, and IX (GnT-Vb). In: Brockhausen, I. (eds) Glycosyltransferases. Methods in Molecular Biology, vol 1022. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-465-4_21
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DOI: https://doi.org/10.1007/978-1-62703-465-4_21
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