Grant M E, Jackson D S
Biochem J. 1968 Jul;108(4):587-91. doi: 10.1042/bj1080587.
Collagen preparations from bovine tissues were analysed for their carbohydrate content. Crude preparations of tropocollagen and polymeric collagen were found to be contaminated with considerable amounts of mannose, fucose and hexosamine, sugars known to be present in the mucoprotein of the interfibrillar material with which collagen is associated in vivo. A pure preparation of tropocollagen obtained by ethanol precipitation procedures contained only galactose and glucose in the approximate ratio of 7:3 residues/3000 amino acid residues. Purification of crude polymeric collagen by EDTA extraction or by crude bacterial amylase extraction considerably decreased the mucoprotein contamination, particularly in the enzymic treatment, which yielded a preparation containing predominantly galactose and glucose in the ratio of 4:2 residues/3000 amino acid residues. The results confirm previous work that demonstrated the purity of these collagen preparations as inferred by amino acid analysis. The results also indicate the suitability of the pure tropocollagen and the amylase-extracted polymeric collagen for studies on the role of the carbohydrate residues in intramolecular and intermolecular cross-linking in collagen.
对源自牛组织的胶原蛋白制剂进行了碳水化合物含量分析。发现原胶原蛋白和聚合胶原蛋白的粗制制剂被大量甘露糖、岩藻糖和己糖胺污染,这些糖存在于体内与胶原蛋白相关的纤维间物质的粘蛋白中。通过乙醇沉淀法获得的原胶原蛋白纯制剂仅含有半乳糖和葡萄糖,其比例约为7:3残基/3000个氨基酸残基。通过EDTA提取或粗制细菌淀粉酶提取对粗制聚合胶原蛋白进行纯化,可显著降低粘蛋白污染,特别是在酶处理中,得到的制剂主要含有半乳糖和葡萄糖,比例为4:2残基/3000个氨基酸残基。结果证实了先前的工作,即通过氨基酸分析推断这些胶原蛋白制剂的纯度。结果还表明,纯原胶原蛋白和经淀粉酶提取的聚合胶原蛋白适用于研究碳水化合物残基在胶原蛋白分子内和分子间交联中的作用。