Lyon M, Deakin J A, Gallagher J T
Cancer Research Campaign, Christie Hospital, Manchester, United Kingdom.
J Biol Chem. 1994 Apr 15;269(15):11208-15.
The structure of rat liver heparan sulfate (HS) has been investigated using a combination of (a) chain scission with specific reagents, (b) disaccharide compositional analysis, and (c) end-referenced sequence analysis of the proximal, protein-linked region of the chain. This study reveals that the liver synthesizes a highly sulfated HS species (1.34 sulfates/disaccharide), particularly high in N-sulfation (60%) and 2-O-sulfate content (36%). Approximately half of the latter is found in trisulfated disaccharides, i.e. IdceA(2-OSO3) alpha 1-4GlcNSO3 (6-OSO3). End-referencing methodology established the existence of an extended, unmodified heparan (GlcUA beta 1-4GlcNAc) sequence, 8-11 disaccharides in length, attached to the linkage tetrasaccharide, similar to that found in a number of other HS species. Directly following this is a mixed HexUA1-4GlcNR(6-OSO3) (where GlcNR represents alpha-D-glucosamine with an unspecified N-substituent)-containing sequence of variable length, culminating in the appearance of the first IdceA(2-OSO3) residue approximately 20 disaccharides from the linkage region, i.e. approximately 40% along the length of the chain. The distal 60% of the polysaccharide is highly sulfated (approximately 2 sulfates/disaccharide) and mainly comprises three heparin-like domains, highly enriched in IdceA(2-OSO3) residues. Overall, liver HS qualifies as an extreme member of the HS family, with a considerable proportion of heparin-like structure asymmetrically concentrated to the distal part of the chain.
利用以下方法组合研究了大鼠肝脏硫酸乙酰肝素(HS)的结构:(a)用特定试剂进行链断裂,(b)二糖组成分析,以及(c)对链近端与蛋白质相连区域进行末端参考序列分析。这项研究表明,肝脏合成了一种高度硫酸化的HS种类(1.34个硫酸盐/二糖),其N-硫酸化(60%)和2-O-硫酸盐含量(36%)特别高。后者约一半存在于三硫酸化二糖中,即艾杜糖醛酸(2-OSO3)α1-4葡糖胺(6-OSO3)。末端参考方法确定了存在一个延伸的、未修饰的乙酰肝素(葡糖醛酸β1-4葡糖胺)序列,长度为8-11个二糖,连接到连接四糖上,类似于在许多其他HS种类中发现的序列。紧接着这个序列的是一个长度可变的混合的HexUA1-4GlcNR(6-OSO3)(其中GlcNR代表具有未指定N-取代基的α-D-葡糖胺)序列,最终在距连接区域约20个二糖处出现第一个艾杜糖醛酸(2-OSO3)残基,即沿链长度约40%处。多糖的远端60%高度硫酸化(约2个硫酸盐/二糖),主要由三个类肝素结构域组成,高度富含艾杜糖醛酸(2-OSO3)残基。总体而言,肝脏HS可被视为HS家族的一个极端成员,相当一部分类肝素结构不对称地集中在链的远端。