Amplification and Preparation of Cellular O-Glycomes for Functional Glycomics.

Analytical Chemistry
Zhonghua LiXuezheng Song

Abstract

Mucin-type O-glycans play key roles in many cellular processes, and they are often altered in human diseases. A major challenge in studying the role of O-glycans through functional O-glycomics is the absence of a complete repertoire of the glycans that comprise the human O-glycome. Here we describe a cellular O-glycome preparation strategy, Preparative Cellular O-Glycome Reporter/Amplification (pCORA), that introduces 4-N3-Bn-GalNAc(Ac)3 as a novel precursor in large-scale cell cultures to generate usable amounts of O-glycans as a potential O-glycome factory. Cultured human non-small cell lung cancer (NSCLC) A549 cells take up the precursor, which is extended by cellular glycosyltransferases to produce 4-N3-Bn-α-O-glycans that are secreted into the culture medium. The O-glycan derivatives can be clicked with a fluorescent bifunctional tag that allows multidimensional HPLC purification and production of a tagged glycan library, representing the O-glycome of the corresponding cells. We obtained ∼5% conversion of precursor to O-glycans and purified a tagged O-glycan library of over 100 O-glycan derivatives, many of which were present in >100 nmol amounts and were sequenced by sequential MS fragmentation (MSn). These O-glycans were...Continue Reading

References

Mar 17, 2000·Science·E Saxon, C R Bertozzi
Dec 6, 2003·Proceedings of the National Academy of Sciences of the United States of America·Howard C HangCarolyn R Bertozzi
Jan 13, 2006·Nature Chemical Biology·Jennifer A Prescher, Carolyn R Bertozzi
Apr 19, 2006·Nature Chemical Biology·James C PaulsonBrian E Collins
May 23, 2007·The Journal of Experimental Medicine·Guangyu AnLijun Xia
Feb 2, 2008·Inflammatory Bowel Diseases·Daniele SimondiMarco Astegiano
Jan 28, 2009·Chemistry & Biology·Xuezheng SongRichard D Cummings
Dec 7, 2010·Nature Methods·Xuezheng SongDavid F Smith
Jan 25, 2011·Angewandte Chemie·Tongzhong JuRichard D Cummings
Feb 26, 2011·International Journal of Cancer. Journal International Du Cancer·Francis JacobViola Heinzelmann-Schwarz
Oct 26, 2011·European Journal of Medical Research·T O HircheOliver Schröder
Apr 16, 2013·The EMBO Journal·Catharina SteentoftHenrik Clausen
Jul 17, 2013·Proteomics. Clinical Applications·Tongzhong JuRichard D Cummings
Dec 18, 2013·Proteomics·Maja N ChristiansenNicolle H Packer
Mar 20, 2014·Cancer Biomarkers : Section a of Disease Markers·Tongzhong JuRichard D Cummings
May 27, 2014·American Society of Clinical Oncology Educational Book·Heather WakeleeMartin J Edelman
Jul 23, 2014·Molecular & Cellular Proteomics : MCP·David J AshlineVernon N Reinhold
Jan 27, 2015·Annual Review of Pathology·Sean R StowellRichard D Cummings
Apr 9, 2015·Frontiers in Genetics·Louise E TailfordNathalie Juge
Dec 1, 2015·Nature Methods·Matthew R KudelkaTongzhong Ju
May 3, 2016·Nature Methods·Xuezheng SongRichard D Cummings
Nov 9, 2016·Analytical and Bioanalytical Chemistry·David J AshlineVernon N Reinhold
Sep 10, 2018·Glycoconjugate Journal·Chengjian WangZhongfu Wang
Nov 2, 2018·Bioconjugate Chemistry·Yuyang ZhuXuezheng Song
Nov 14, 2018·Translational Research : the Journal of Laboratory and Clinical Medicine·Mariska DavidsMay Christine V Malicdan
Jan 11, 2019·Biochemical Society Transactions·David F SmithXuezheng Song
Jul 1, 2019·Methods in Molecular Biology·Detlef Grunow, Véronique Blanchard

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