Cheng F, Havsmark B, Sakurai K, Habuchi H, Suzuki S, Yoshida K, Fransson L A
Department of Cell and Molecular Biology, Lund University, Sweden.
Glycoconj J. 1997 Feb;14(2):297-305. doi: 10.1023/a:1018562426363.
By supplying skin fibroblasts with different concentrations of the galactosaminoglycan chain-primer p-hydroxyphenyl-O-beta-D-xylopyranoside we have produced and recovered glycan-chains that were subsequently radio-iodinated in the hydroxyphenyl group and subjected to sequence analysis by using graded enzymic treatment followed by a combination of gel chromatography and electrophoresis. Fragments extending from the tagged reducing end to the cleavage-point were identified and quantified. Degradation by chondroitin B lyase of chains primed at 0.1 or 0.5 mM xyloside gave profiles indicating a periodic and wave-like distribution of iduronate-containing repeats, with high incidence around positions 2, 5 and onwards, whereas in chains produced at 1.0 mM xyloside the incidence of iduronate was similar in positions 1-4 and then declined. Degradation by chondroitin AC lyase indicated a high incidence of glucuronate in or near the linkage-region. There was a relatively uniform degree of sulfation in chains primed at low xyloside concentration, whereas chains primed at 1.0 mM xyloside gave very heterogeneous charge-patterns in all segments of the chain, including the linkage-region, giving the impression that adequate sulfation, probably at C-4 and at the first opportunity, is necessary to obtain an ordered and periodic epimerization pattern.
通过向皮肤成纤维细胞提供不同浓度的半乳糖胺聚糖链引物对羟基苯基 - O - β - D - 木吡喃糖苷，我们制备并回收了聚糖链，随后在羟基苯基上进行放射性碘化，并通过分级酶处理，然后结合凝胶色谱和电泳进行序列分析。鉴定并定量了从标记的还原端延伸至切割点的片段。用软骨素B裂解酶对以0.1或0.5 mM木糖苷引发的链进行降解，得到的图谱表明含艾杜糖醛酸的重复序列呈周期性和波浪状分布，在位置2、5及之后出现的频率较高，而在以1.0 mM木糖苷产生的链中，艾杜糖醛酸在位置1 - 4的出现频率相似，然后下降。用软骨素AC裂解酶降解表明在连接区或其附近葡糖醛酸的出现频率较高。在低木糖苷浓度引发的链中硫酸化程度相对均匀，而在以1.0 mM木糖苷引发的链中，在链的所有片段（包括连接区）中电荷模式非常不均一，这给人的印象是，可能在C - 4且在第一个机会时进行充分的硫酸化对于获得有序和周期性差向异构化模式是必要的。
