title: Simultaneous analysis of heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan disaccharides by glycoblotting-assisted sample preparation followed by single-step zwitter-ionic-hydrophilic interaction chromatography.
abstract: Glycosaminoglycans (GAGs) play important roles in cell adhesion and growth, maintenance of extracellular matrix (ECM) integrity, and signal transduction. To fully understand the biological functions of GAGs, there is a growing need for sensitive, rapid, and quantitative analysis of GAGs. The present work describes a novel analytical technique that enables high throughput cellular/tissue glycosaminoglycomics for all three families of uronic acid-containing GAGs, hyaluronan (HA), chondroitin sulfate (CS)/dermatan sulfate (DS), and heparan sulfate (HS). A one-pot purification and labeling procedure for GAG Δ-disaccharides was established by chemo-selective ligation of disaccharides onto high density hydrazide beads (glycoblotting) and subsequent labeling by fluorescence. The 17 most common disaccharides (eight comprising HS, eight CS/DS, and one comprising HA) could be separated with a single chromatography for the first time by employing a zwitter-ionic type of hydrophilic-interaction chromatography column. These novel analytical techniques were able to precisely characterize the glycosaminoglycome in various cell types including embryonal carcinoma cells and ocular epithelial tissues (cornea, conjunctiva, and limbus).
authors: Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, Sapporo, Japan.
authors: Yasuhiro
authors: Y
authors: Takegawa
authors: Kayo
authors: K
authors: Araki
authors: Naoki
authors: N
authors: Fujitani
authors: Jun-ichi
authors: J
authors: Furukawa
authors: Hiroaki
authors: H
authors: Sugiyama
authors: Hideaki
authors: H
authors: Sakai
authors: Yasuro
authors: Y
authors: Shinohara
mesh_terms: D000818:Animals; D002460:Cell Line; D002809:Chondroitin Sulfates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D003228:Conjunctiva / Q000378:metabolism; D006224:Cricetinae; D003871:Dermatan Sulfate / Q000032:analysis*; D004187:Disaccharides / Q000032:analysis; D006497:Heparan Sulfate / Q000032:analysis*; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis*; D057927:Hydrophobic and Hydrophilic Interactions; D007477:Ions / Q000737:chemistry; D051379:Mice; D011817:Rabbits
chemical_list: D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D004187:Disaccharides; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D007477:Ions