Turpeenniemi T M, Puistola U, Anttinen H, Kivirikko K I
Biochim Biophys Acta. 1977 Jul 8;483(1):215-9. doi: 10.1016/0005-2744(77)90024-9.
Lysyl hydroxylase (peptidyllysine, 2-oxoglutarate: oxygen 5-oxidoreductase, EC 1.14.11.4) has a high affinity for columns of concanavalin A-agarose, which was markedly reduced in the presence of alpha-methyl-D-mannoside, suggesting that the enzyme is a glycoprotein. Once bound, the enzyme could not be eluted with the glycoside alone, whereas an effective elution was achieved by a combination of alpha-methyl-D-mannoside and ethylene glycol. The data thus suggest that hydrophobic interaction stabilized the complex of the enzyme with the column. This information was applied to obtain a lysyl hydroxylase purification of about 3000-fold with a recovery of more than 10% from extract of chick embryos by relatively simple steps.
赖氨酰羟化酶（肽基赖氨酸，2-氧代戊二酸：氧5-氧化还原酶，EC 1.14.11.4）对伴刀豆球蛋白A-琼脂糖柱具有高亲和力，在α-甲基-D-甘露糖苷存在下其亲和力显著降低，这表明该酶是一种糖蛋白。一旦结合，仅用糖苷无法洗脱该酶，而通过α-甲基-D-甘露糖苷和乙二醇的组合可实现有效洗脱。因此，数据表明疏水相互作用稳定了酶与柱的复合物。通过相对简单的步骤，利用该信息从鸡胚提取物中获得了约3000倍的赖氨酰羟化酶纯化，回收率超过10%。
