Uchiyama H, Nagasawa K
Carbohydr Res. 1987 Feb 1;159(2):263-73. doi: 10.1016/s0008-6215(00)90220-9.
A method for analyzing the distribution of D-glucuronic acid units within the chain and near the linkage region of dermatan sulfate has been developed. The method consists of a chemical modification of the reducing terminal residue in the polysaccharide by reductive amination with excess 1,2-diaminoethane in the presence of sodium cyanoborohydride, desulfative fragmentation of the polysaccharide, labeled with 2-aminoethylamino (AEA) groups, in hot dimethyl sulfoxide containing 10% of water followed by 2,4-dinitrophenylation of the 2-aminoethylamino group, separation of the 2-(2,4-dinitrophenylamino)ethylamino labeled dermatan fragments from nonlabeled fragments on Octyl-Sepharose CL-4B gel, and determination of the uronic acid composition of the labeled fragments having various chain-length. A preparation of pig-skin dermatan sulfate (Mr 21,000, ratio of GlcA to total uronic acid, 93:500) showed an average distribution pattern of D-glucuronic acid residues near the linkage region of one N-acetylchondrosine unit in the disaccharide sequence 1-5(6) linked to the Xyl----Gal----Gal----GlcA residue, a cluster of 6-8 N-acetyldermosine units in the sequence 6(7)-12(13), and four separate N-acetylchondrosine units between the sequence adjacent to the N-acetyldermosine cluster and the sequence 23 or higher.
已开发出一种分析硫酸皮肤素链内及连接区域附近D-葡萄糖醛酸单元分布的方法。该方法包括：在氰基硼氢化钠存在下，用过量的1,2-二氨基乙烷对多糖的还原末端残基进行还原胺化化学修饰；在含10%水的热二甲基亚砜中，对用2-氨基乙氨基（AEA）基团标记的多糖进行脱硫断裂；然后对2-氨基乙氨基进行2,4-二硝基苯化；在辛基-琼脂糖CL-4B凝胶上，将2-(2,4-二硝基苯氨基)乙氨基标记的硫酸皮肤素片段与未标记片段分离；并测定具有不同链长的标记片段的糖醛酸组成。一种猪皮硫酸皮肤素制剂（Mr 21,000，葡萄糖醛酸与总糖醛酸的比例为93:500）显示，在与木糖-半乳糖-半乳糖-葡萄糖醛酸残基相连的二糖序列1-5(6)中，一个N-乙酰软骨素单元连接区域附近的D-葡萄糖醛酸残基呈平均分布模式；在序列6(7)-12(13)中有一簇6-8个N-乙酰皮肤素单元；在与N-乙酰皮肤素簇相邻的序列和序列23或更高序列之间有四个独立的N-乙酰软骨素单元。
