Comprehensive approach to structural and functional glycomics based on chemoselective glycoblotting and sequential tag conversion

Analytical Chemistry
Jun-ichi FurukawaShin-Ichiro Nishimura

Abstract

Changes in protein glycosylation profoundly affect protein function. To understand these effects of altered protein glycosylation, we urgently need high-throughput technologies to analyze glycan expression and glycan-protein interactions. Methods are not available for amplification of glycans; therefore, highly efficient sample preparation is a major issue. Here we present a novel strategy that allows flexible and sequential incorporation of various functional tags into oligosaccharides derived from biological samples in a practical manner. When combined with a chemoselective glycoblotting platform, our analysis enables us to complete sample preparation (from serum to released, purified, methyl-esterified, and labeled glycans) in 8 h from multiple serum samples (up to 96 samples) using a 96-well microplate format and a standard de-N-glycosylation protocol that requires reductive alkylation and tryptic digestion prior to PNGase F digestion to ensure maximal de-N-glycosylation efficiency. Using this technique, we quantitatively detected more than 120 glycans on human carcinoembryonic antigens for the first time. This approach was further developed to include a streamlined method of purification, chromatographic fractionation, and...Continue Reading

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Citations

Mar 13, 2009·Journal of the American Society for Mass Spectrometry·Kathryn R RebecchiHeather Desaire
Jun 24, 2010·Molecular & Cellular Proteomics : MCP·Masaki KurogochiShin-Ichiro Nishimura
May 7, 2010·Omics : a Journal of Integrative Biology·Joseph Zaia
Oct 2, 2012·Journal of the American Society of Nephrology : JASN·Masako OtaniShozo Izui
Jan 25, 2013·Proceedings of the National Academy of Sciences of the United States of America·Naoki FujitaniYasuro Shinohara
Apr 24, 2014·International Journal of Molecular Sciences·Zining CuiYoshihiro Nishida
Feb 9, 2010·Bioscience, Biotechnology, and Biochemistry·Mizuho InagakiTohru Suzuki
Aug 21, 2014·Proceedings of the National Academy of Sciences of the United States of America·Yanlu CaoC Jeffrey Smith
Mar 9, 2011·Journal of Pharmaceutical and Biomedical Analysis·Keita Yamada, Kazuaki Kakehi
Jul 25, 2009·Current Opinion in Chemical Biology·Oyindasola Oyelaran, Jeffrey C Gildersleeve
Aug 5, 2015·Glycoconjugate Journal·L S Mamatha BhanuH S Aparna
Oct 24, 2012·Proteomics. Clinical Applications·Shuang Yang, Hui Zhang
Jan 17, 2013·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Toshiya KamiyamaSatoru Todo
Aug 13, 2011·The FEBS Journal·Shota TakimoriShin-Ichiro Nishimura
May 31, 2015·Plant Science : an International Journal of Experimental Plant Biology·Sho TakanoKiyoaki Kato
Jul 2, 2015·Analytical Chemistry·Jun-ichi FurukawaYasuro Shinohara
Apr 25, 2015·Biochimica Et Biophysica Acta·Solomon T GizawShin-Ichiro Nishimura
Jul 14, 2012·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Naomi ManriTakeshi Sakamoto
Mar 31, 2015·Chromatographia·Archana ShubhakarManfred Wuhrer
Jun 21, 2016·Transplantation Proceedings·H EguchiS Miyagawa
Oct 29, 2015·Chemical Reviews·Yoshiko MiuraHirokazu Seto
Nov 4, 2016·Proteomics·Yasunobu YoshidaYasuro Shinohara
Feb 2, 2017·Journal of Agricultural and Food Chemistry·Nagaraju BrijeshaHuligerepura Sosalegowda Aparna
Oct 1, 2008·Molecular & Cellular Proteomics : MCP·Rie UematsuShin-ichiro Nishimura
Oct 1, 2015·Journal of Lipid Research·Jun-ichi FurukawaYasuro Shinohara
Jul 21, 2015·Integrative Biology : Quantitative Biosciences From Nano to Macro·H StöckmannP M Rudd
Jun 20, 2017·Journal of Proteome Research·Jun-Ichi FurukawaYasuro Shinohara

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