Characterizing the Microstructure of Heparin and Heparan Sulfate Using N-Sulfoglucosamine 1H and 15N NMR Chemical Shift Analysis

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• 作者：Derek J. Langeslay ; Consuelo N. Beecher ; Annamaria Naggi ; Marco Guerrini ; Giangiacomo Torri ; Cynthia K. Larive
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：January 15, 2013
• 年：2013
• 卷：85
• 期：2
• 页码：1247-1255
• 全文大小：314K
• 年卷期：v.85,no.2(January 15, 2013)
• ISSN：1520-6882

文摘

Heparin and heparan sulfate (HS) are members of a biologically important group of highly anionic linear polysaccharides called glycosaminoglycans (GAGs). Because of their structural complexity, the molecular-level characterization of heparin and HS continues to be a challenge. The work presented herein describes an emerging approach for the analysis of unfractionated and low molecular weight heparins, as well as porcine and human-derived HS. This approach utilizes the untapped potential of ¹⁵N NMR to characterize these preparations through detection of the NH resonances of N-sulfo-glucosamine residues. The sulfamate group ¹H and ¹⁵N chemical shifts of six GAG microenvironments were assigned based on the critical comparison of selectively modified heparin derivatives, NMR measurements for a library of heparin-derived oligosaccharide standards, and an in-depth NMR analysis of the low molecular weight heparin enoxaparin through systematic investigation of the chemical exchange properties of NH resonances and residue-specific assignments using the [¹H,¹⁵N] HSQC-TOCSY experiment. The sulfamate microenvironments characterized in this study include GlcNS(6S)-UA(2S), 螖UA(2S)-GlcNS(6S), GlcNS(3S)(6S)-UA(2S), GlcNS-UA, GlcNS(6S)-red_伪, and 1,6-anhydro GlcNS demonstrating the utility of [¹H,¹⁵N] HSQC NMR spectra to provide a spectroscopic fingerprint reflecting the composition of intact GAGs and low molecular weight heparin preparations.