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: NMR Spectroscopy The sample (30 mg) was dissolved in deuterium oxide (99.9 % D, Aldrich) and lyophilized. The sample was dissolved in 96 ?L D2O (99.96 % D, Cambridge Isotope), 4 ?L 50 mg/mL DSS was added, and the solution placed in a 3-mm Shigemi NMR tube. 1-D Proton, gCOSY, TOCSY, gHSQC, NOESY, and gHMBC NMR spectra were acquired on a Varian Inova-500 spectrometer at 298 K (25 ?C). Chemical shifts were measured relative to internal DSS (?H=0.000 ppm, ?C=0.000 ppm). Further experimental details are available upon request. ESI Mass Spectrometry Mass spectrometry was performed on an LCQ Advantage ESI-quadrupole instrument using 50% MeOH/water as spray solvent. The instrument was tuned on the trisulfated heparin disaccharide I-S. Spray voltage was 3.80 kV, capillary temperature 200 ?C, sheath gas flow rate 43, capillary voltage -11.00 V, and tube lens offset 15.00 V. A ~50 ?M solution was infused via syringe pump and silica capillary at 10 ?L/min. Signal averaging was performed over 2 min scan time.
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