This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Methods: Preparation of Partially Methylated Alditol Acetates (PMAAs) For the determination of linked glycosyls of that composed the N-linked glycans, PMAAs were prepared from the permethylated oligosaccharides (remainder from oligosaccharide profiling by MALDI-TOF/TOF MS). Briefly, permethylated oligosaccharides were hydrolyzed with HCL:water:acetic acid (1:3:16) at 80oC overnight, followed by reduction with NaBD4. The latter-freed hydroxyls after hydrolysis were acetylated with acetic anhydride/pyridine (1:1, v/v) at 100 ?C for 1 h. Gas Chromatograph-Mass Spectrometry (GC-MS) The PMAAs for amino sugars were analyzed on a Hewlett Packard 5890 GC interfaced to a 5970 MSD, which was installed with a 30-m EC 1 bonded phase fused silica capillary column (Altech), whereas the neutral sugars were analyzed on an Agilent 7890A GC interfaced to a 5975C MSD equipped with a Rtx-2330 column. The GC conditions were the following: For amino sugars ? initial oven temperature was 140?C (2.0?C/min), which was held for 0.5 min, ramped to 220?C (20?C/min) and finally ramped to 300?C (20?C/min) and held for 7.5 min. For neutral sugars ? initial oven temp was 80oC, which was held for 2 min, ramped to 170oC at 30oC/min, held for 0 min, and finally ramped to 235oC at 4oC/min and held for 20 min
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