Nelson S R, Lyon M, Gallagher J T, Johnson E A, Pepys M B
Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.
Biochem J. 1991 Apr 1;275 ( Pt 1)(Pt 1):67-73. doi: 10.1042/bj2750067.
Amyloid fibrils were isolated by extraction in water from the livers and spleens of four patients who had died of monoclonal, light-chain (AL)-type, systemic amyloidosis and one with reactive systemic, amyloid A protein (AA)-type amyloidosis. Each fibril preparation contained 1-2% by weight of glycosaminoglycan (GAG) which was tightly associated with the fibrils and not just co-isolated from the tissues with them. After exhaustive digestion of the fibrils with papain and Pronase, the GAGs were specifically precipitated with cetylpyridinium chloride and were identified by cellulose acetate electrophoresis and selective susceptibility to specific glycosidases. All the preparations contained approximately equal amounts of heparan sulphate and dermatan sulphate. There was no evidence for the presence of chondroitin sulphate or other GAGs. Fine structural analysis by oligosaccharide mapping in gradient polyacrylamide gels, following partial digestion with specific glycosidases, showed very similar structures among the heparan sulphates and the dermatan sulphates, respectively. GAGs were also extracted by solubilizing amyloid fibrils in 4 M-guanidinium chloride followed by CsCl density-gradient ultracentrifugation. Although a minor proportion of the GAG material obtained in this way was apparently in the form of proteoglycan molecules, most of it was free GAG chains. The presence in amyloid fibrils of different types, in different organs and from different patients of particular GAG classes with similar structures supports the view that these molecules may be of pathogenic significance.
从死于单克隆轻链（AL）型系统性淀粉样变性的4例患者以及1例反应性系统性淀粉样A蛋白（AA）型淀粉样变性患者的肝脏和脾脏中，通过水提取法分离出淀粉样纤维。每种纤维制剂含有1-2%（重量）的糖胺聚糖（GAG），其与纤维紧密结合，并非仅仅是与它们从组织中共同分离出来。用木瓜蛋白酶和链霉蛋白酶彻底消化纤维后，GAG用十六烷基氯化吡啶特异性沉淀，并通过醋酸纤维素电泳和对特定糖苷酶的选择性敏感性进行鉴定。所有制剂都含有大致等量的硫酸乙酰肝素和硫酸皮肤素。没有证据表明存在硫酸软骨素或其他GAG。在用特定糖苷酶部分消化后，通过梯度聚丙烯酰胺凝胶中的寡糖图谱进行精细结构分析，结果显示硫酸乙酰肝素和硫酸皮肤素之间的结构非常相似。还通过将淀粉样纤维溶解在4M盐酸胍中，然后进行CsCl密度梯度超速离心来提取GAG。虽然以这种方式获得的一小部分GAG物质显然是以蛋白聚糖分子的形式存在，但大部分是游离的GAG链。不同类型、不同器官以及不同患者的淀粉样纤维中存在结构相似的特定GAG类别，这支持了这些分子可能具有致病意义的观点。
