Fransson L A, Huckerby T N, Nieduszynski I A
Biochem J. 1978 Oct 1;175(1):299-309. doi: 10.1042/bj1750299.
A heparin derivative that had been O/N-desulphated and re-N-acetylated was investigated by 13C n.m.r. spectroscopy and potentiometric titration. Three forms of uronic acid were observed, tentatively identified as beta-D-glucuronate, and two different forms of alpha-L-iduronate. A comparison of the n.m.r. spectra of heparin, an oligosaccharide (beta-D-glucuronate-2-acetamido-2-deoxy-alpha-D-glucose)n, and heparin that had been subjected to selective oxidation of beta-D-glucuronate, enabled the position of the anomeric carbon of the latter residue to be assigned [delta 102.9 (p.p.m.)]. Periodate oxidation of O/N-desulphated heparin destroyed in addition, approx. 40% of the alpha-L-iduronate content. The remainder of the alpha-L-iduronate residues displayed only one anomeric resonance, at delta 99.7 (p.p.m.). In another preparation, after sequential desulphation of heparin (N-desulphation, re-N-acetylation and O-desulphation) the anomeric resonance of the alpha-L-iduronate residue shifted downfield [from delta99.7 (p.p.m.) to delta 102.3]indicating a change in ring conformation. These data support the interpretation that the unsulphated alpha-L-iduronate residues may adopt two conformations. It was shown that the proportions of alpha-L-iduronate conformers are determined by the sequence of desulphation operations. Also minor components of heparin were assigned.
通过(^{13}C)核磁共振光谱和电位滴定法研究了一种经过过夜脱硫和重新(N)-乙酰化的肝素衍生物。观察到三种形式的糖醛酸,初步鉴定为β-D-葡萄糖醛酸,以及两种不同形式的α-L-艾杜糖醛酸。通过比较肝素、寡糖(β-D-葡萄糖醛酸-2-乙酰氨基-2-脱氧-α-D-葡萄糖)(n)和经过β-D-葡萄糖醛酸选择性氧化的肝素的核磁共振光谱,确定了后一种残基的异头碳位置[δ102.9(ppm)]。此外,对经过(O/N)-脱硫的肝素进行高碘酸盐氧化,破坏了约40%的α-L-艾杜糖醛酸含量。其余的α-L-艾杜糖醛酸残基仅显示一个异头共振峰,位于δ99.7(ppm)。在另一种制备方法中,对肝素进行连续脱硫((N)-脱硫、重新(N)-乙酰化和(O)-脱硫)后,α-L-艾杜糖醛酸残基的异头共振峰向低场移动[从δ99.7(ppm)移至δ102.3],表明环构象发生了变化。这些数据支持了未硫酸化的α-L-艾杜糖醛酸残基可能采用两种构象的解释。结果表明,α-L-艾杜糖醛酸构象异构体的比例由脱硫操作顺序决定。同时还确定了肝素的次要成分。