Huckerby T N, Dickenson J M, Brown G M, Nieduszynski I A
Polymer Centre, Lancaster University, UK.
Biochim Biophys Acta. 1995 May 11;1244(1):17-29. doi: 10.1016/0304-4165(94)00180-6.
Keratan sulfate (KS) chains prepared from both bovine tracheal rings and bovine femoral head cartilage were digested with the enzyme keratanase from Pseudomonas species; large repeat-sequence and non-reducing terminal oligosaccharides were fractionated and purified using high-performance ion-exchange chromatography. The main beta-linked pentasulfated hexasaccharide repeat segment, [R6], GlcNAc(6S)1-1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal-ol and the asialo beta-linked capping pentasulfated heptasaccharide, [C7], Gal1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S) 1-3Gal-ol have been completely characterized by high-field NMR spectroscopy using one- and two-dimensional methods. Partial 1H assignments are summarized for three homologous series of higher oligosaccharides: GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-5(1-3)Gal-0l [R8,R10,R12] Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]3-5(1-3)Gal-ol [C9,C11,C13] NeuAc alpha 2-3Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-4(1-3)Gal-ol [C8,C10,C12] obtained from keratan sulfate by keratanase cleavage. The first shows that the unsulfated galactose residues within the repeat sequence region of KS may be separated by fully sulfated segments which have a wide distribution of lengths. The others, viz. those with sialylated caps, and the related galactose capped asialo-segments (derived from a KS digestion in which the keratanase also exhibited sialidase activity) represent an homologous series of epitopes in which the first internal unsulfated galactose is located at a position which may be up to five or more fully sulfated N-acetyllactosamine disaccharide repeat units along from the non-reducing terminus of the KS polymer.
从牛气管环和牛股骨头软骨中制备的硫酸角质素（KS）链，用假单胞菌属的角质素酶进行消化；使用高效离子交换色谱法对大的重复序列和非还原末端寡糖进行分级分离和纯化。主要的β-连接的五硫酸化六糖重复片段，[R6]，GlcNAc(6S)1-1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal-ol和去唾液酸β-连接的封端五硫酸化七糖，[C7]，Gal1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S) 1-3Gal-ol已通过使用一维和二维方法的高场核磁共振光谱法进行了完全表征。总结了三个同源系列的高级寡糖的部分1H归属：GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-5(1-3)Gal-0l [R8,R10,R12] Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]3-5(1-3)Gal-ol [C9,C11,C13] NeuAcα2-3Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-4(1-3)Gal-ol [C8,C10,C12]，这些寡糖是通过角质素酶切割硫酸角质素获得的。第一个结果表明，KS重复序列区域内的非硫酸化半乳糖残基可能被长度分布广泛的完全硫酸化片段隔开。其他的，即那些带有唾液酸化帽的，以及相关的半乳糖封端的去唾液酸片段（源自角质素酶也表现出唾液酸酶活性的KS消化）代表了一个同源系列的表位，其中第一个内部非硫酸化半乳糖位于一个位置，该位置可能距离KS聚合物的非还原末端多达五个或更多个完全硫酸化的N-乙酰乳糖胺二糖重复单元。
