Schmidt A, Schäfer E, Buddecke E
Institut für Arterioskleroseforschung, Münster, Federal Republic of Germany.
Eur J Biochem. 1988 May 2;173(3):661-6. doi: 10.1111/j.1432-1033.1988.tb14049.x.
When calf aortic tissue, preincubated under organ culture conditions in the presence of [35S]sulfate, was submitted to a sequential collagenase and elastase digestion and guanidinium chloride extraction, the bulk of proteoheparan sulfate was obtained in the elastase fraction. Ion-exchange chromatography on DEAE-cellulose of the elastase digest under dissociative conditions yielded a proteoglycan fraction that contained heparan sulfate as the sole glycosaminoglycan. The proteoheparan sulfate fraction was resolved into a high-molecular-mass (P-HS 1) and a low-molecular-mass (P-HS 2) fraction by gel filtration on Sephacryl S-400. P-HS 1 has a Mr of 175,000 and possesses four heparan sulfate side-chains (Mr 32,000) covalently bound to the protein core via a galactose- and xylose-containing polysaccharide-protein binding region. The protein core (Mr 38,000), which was obtained after deglycosylation of PG-HS 1 with trifluormethane sulfonic acid, contained in addition a few N-glycosidically linked oligosaccharide units representing a complex type with terminal neuraminic acid residues. P-HS 2 is a single-chain peptidoheparan sulfate of Mr of 38,000 containing one heparan sulfate chain (Mr 32,000) linked to a polypeptide (Mr 6000). The ratio of specific radioactivities of P-HS 1 and P-HS 2 was 1:0.66.
当在器官培养条件下于[35S]硫酸盐存在的情况下预孵育的小牛主动脉组织，经过胶原酶和弹性蛋白酶的顺序消化以及氯化胍提取后，大部分蛋白聚糖硫酸乙酰肝素在弹性蛋白酶部分中获得。在解离条件下对弹性蛋白酶消化产物进行DEAE - 纤维素离子交换色谱分析，得到一个蛋白聚糖部分，其中硫酸乙酰肝素是唯一的糖胺聚糖。通过在Sephacryl S - 400上进行凝胶过滤，将蛋白聚糖硫酸乙酰肝素部分分离为高分子量（P - HS 1）和低分子量（P - HS 2）部分。P - HS 1的分子量为175,000，具有四个硫酸乙酰肝素侧链（分子量32,000），通过含半乳糖和木糖的多糖 - 蛋白质结合区域与蛋白质核心共价结合。在用三氟甲磺酸对PG - HS 1进行去糖基化后得到的蛋白质核心（分子量38,000），还含有一些N - 糖苷键连接的寡糖单元，代表一种带有末端神经氨酸残基的复杂类型。P - HS 2是一种分子量为38,000的单链肽聚糖硫酸乙酰肝素，含有一条硫酸乙酰肝素链（分子量32,000）与一条多肽（分子量6000）相连。P - HS 1和P - HS 2的比放射性之比为1:0.66。
