Furlan M, Perret B A, Beck E A
Schweiz Med Wochenschr. 1979 Sep 29;109(37):1369-70.
Human and bovine factor VIII, isolated from cryoprecipitates of fresh plasma by gel filtration on Sepharose CL-2B, gave similar elution patterns and showed comparable distribution of oligomers on SDS agarose electrophoretic gels. The carbohydrate content of individual factor VIII bands, measured by reaction with dansyl hydrazine or binding of glucose/mannose specific concanavalin A, was not directly related to the size or von Willebrand activity of factor VIII oligomers. Staining of disulfide-reduced factor VIII subunits, in polyacrylamide gels, with galactose-specific fluorescein-labelled Ricinus communis lectins, showed an increased binding affinity with increasing size and von Willebrand activity of the parent factor VIII. The von Willebrand activity was strongly inhibited by reaction with Ricinus RCAI lectin, whereas concanavalin A inhibited platelet aggregation only at concentrations above 1 mg/ml. These results suggest that galactose residues are involved in the aggregation of platelets by factor VIII.
通过在Sepharose CL - 2B上进行凝胶过滤从新鲜血浆冷沉淀中分离出的人源和牛源凝血因子VIII，呈现出相似的洗脱模式，并且在SDS琼脂糖电泳凝胶上显示出可比的寡聚体分布。通过与丹磺酰肼反应或结合葡萄糖/甘露糖特异性伴刀豆球蛋白A测量的各个凝血因子VIII条带的碳水化合物含量，与凝血因子VIII寡聚体的大小或血管性血友病活性没有直接关系。在聚丙烯酰胺凝胶中，用半乳糖特异性荧光素标记的蓖麻凝集素对二硫键还原的凝血因子VIII亚基进行染色，结果显示随着亲本凝血因子VIII大小和血管性血友病活性的增加，结合亲和力增强。血管性血友病活性通过与蓖麻RCAI凝集素反应而受到强烈抑制，而伴刀豆球蛋白A仅在浓度高于1 mg/ml时才抑制血小板聚集。这些结果表明半乳糖残基参与了凝血因子VIII介导的血小板聚集。
