Phosphorylation of uridine and cytidine nucleoside analogs by two human uridine-cytidine kinases
Phosphorylation of uridine and cytidine nucleoside analogs by two human uridine-cytidine kinases
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DOI：
10.1124/mol.59.5.1181
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发表时间：
2001-05-01
影响因子：
3.6
通讯作者：
Karlsson, A
中科院分区：
文献类型：
--
作者：
Van Rompay, AR;Norda, A;Karlsson, A
尿苷-胞苷激酶(UCK)对于核苷类似物的磷酸化具有重要作用，目前正在研究核苷类似物在癌症化疗中的可能用途。我们已经克隆了两个人乳房的cdna。在大肠杆菌中表达了约30 kDa的UCK1和UCK2蛋白，并证明它们能催化URD和CyD的磷酸化。这些酶不能磷酸化脱氧核糖核苷或嘌呤核糖核苷。UCK1mRNA被检测到约1.8和约2.7kb两种异构体。2.7kb的条带在被调查的组织中普遍表达。约1.8kb的条带存在于骨骼肌、心脏、肝脏和肾脏中。仅在胎盘组织中检测到1.2和2.0kb两种UCK2mRNA亚型。编码UCK1和UCK2的基因分别定位在染色体9q34.2-9q34.3和1q22-1q23.2。我们测试了28个胞苷和尿苷核苷类似物作为酶的可能底物。这些酶可磷酸化6-氮卓啶、5-氟尿苷、4-硫代尿苷、5-溴代尿苷、N-4-乙酰胞苷、N-4-苯甲酰胞苷、5-氟胞苷、2-硫代胞苷、5-甲基胞苷和N-4-山梨基胞苷等类似物。这两个人UCK的克隆和重组表达对于开发新型的嘧啶核糖核苷类似物及其药理活性鉴定具有重要意义。
Uridine-cytidine kinases (UCK) have important roles for the phosphorylation of nucleoside analogs that are being investigated for possible use in chemotherapy of cancer. We have cloned the cDNA of two human UCKs. The approximate to 30- kDa proteins, named UCK1 and UCK2, were expressed in Escherichia coli and shown to catalyze the phosphorylation of Urd and Cyd. The enzymes did not phosphorylate deoxyribonucleosides or purine ribonucleosides. UCK1 mRNA was detected as two isoforms of approximate to1.8 and approximate to2.7 kb. The 2.7-kb band was ubiquitously expressed in the investigated tissues. The band of approximate to1.8 kb was present in skeletal muscle, heart, liver, and kidney. The two isoforms of UCK2 mRNA of 1.2 and 2.0 kb were only detected in placenta among the investigated tissues. The genes encoding UCK1 and UCK2 were mapped to chromosome 9q34.2-9q34.3 and 1q22-1q23.2, respectively. We tested 28 cytidine and uridine nucleoside analogs as possible substrates of the enzymes. The enzymes phosphorylated several of the analogs, such as 6-azauridine, 5-fluorouridine, 4-thiouridine, 5-bromouridine, N-4-acetylcytidine, N-4-benzoylcytidine, 5-fluorocytidine, 2-thiocytidine, 5-methylcytidine, and N-4-anisoylcytidine. The cloning and recombinant expression of the two human UCKs will be important for development of novel pyrimidine ribonucleoside analogs and the characterization of their pharmacological activation.