Gan T E, Brumley J L, Van der Weyden M B
J Biol Chem. 1983 Jun 10;258(11):7000-4.
Cytosolic thymidine kinase (EC 2.7.1.21) has been purified 5200-fold to apparent homogeneity from normal human placenta. The purification includes sequential affinity chromatography on blue-Sepharose and a thymidine column. The molecular weight of the enzyme determined by gel filtration and sucrose density ultracentrifugation is 92,000. The subunit molecular weight is 44,000, suggesting that the enzyme is a dimer in its native state. With isoelectric focusing, placental thymidine kinase demonstrated a single form with an isoelectric point of 9.1. The final purified enzyme preparation exhibits no immunological cross-reactivity with human mitochondrial thymidine kinase.
已从正常人胎盘中将胞质胸苷激酶(EC 2.7.1.21)纯化了5200倍,达到表观均一性。纯化过程包括依次在蓝色琼脂糖凝胶和胸苷柱上进行亲和层析。通过凝胶过滤和蔗糖密度超速离心法测定,该酶的分子量为92,000。亚基分子量为44,000,表明该酶在天然状态下是二聚体。经等电聚焦分析,胎盘胸苷激酶呈现单一形式,等电点为9.1。最终纯化的酶制剂与人线粒体胸苷激酶无免疫交叉反应。