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Cloning and expression of human deoxyguanosine kinase cDNA.

机译:人脱氧鸟苷激酶激酶cDNA的克隆和表达。

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摘要

A human cDNA sequence homologous to human deoxycytidine kinase (dCK; EC 2.7.1.74) was identified in the GenBank sequence data base. The longest open reading frame encoded a protein that was 48% identical to dCK at the amino acid level. The cDNA was expressed in Escherichia coli and shown to encode a protein with the same substrate specificity as described for the mitochondrial deoxyguanosine kinase (dGK; EC 2.7.1.113). The N terminus of the deduced amino acid sequence had properties characteristic for a mitochondrial translocation signal, and cleavage at a putative mitochondrial peptidase cleavage site would give a mature protein size of 28 kDa. Northern blot analysis determined the length of dGK mRNA to 1.3 kbp with no cross-hybridization to the 2.8-kbp dCK mRNA. dGK mRNA was detected in all tissues investigated with the highest expression levels in muscle, brain, liver, and lymphoid tissues. Alignment of the dGK and herpes simplex virus type 1 thymidine kinase amino acid sequences showed that five regions, including the substrate-binding pocket and the ATP-binding glycine loop, were also conserved in dGK. To our knowledge, this is the first report of a cloned mitochondrial nucleoside kinase and the first demonstration of a general sequence homology between two mammalian deoxyribonucleoside kinases. Our findings suggest that dCK and dGK are evolutionarily related, as well as related to the family of herpes virus thymidine kinases.
机译:在GenBank序列数据库中鉴定了与人脱氧胞苷激酶(dCK; EC 2.7.1.74)同源的人cDNA序列。最长的开放阅读框编码的蛋白质在氨基酸水平上与dCK具有48%的同一性。 cDNA在大肠杆菌中表达,显示编码的蛋白质具有与针对线粒体脱氧鸟苷激酶(dGK; EC 2.7.1.113)所述相同的底物特异性。推导的氨基酸序列的N末端具有线粒体易位信号的特性,并且在假定的线粒体肽酶裂解位点裂解将产生28kDa的成熟蛋白大小。 Northern印迹分析确定了dGK mRNA的长度至1.3kbp,而没有与2.8kbp dCK mRNA的交叉杂交。在所有调查的组织中均检测到dGK mRNA,在肌肉,大脑,肝脏和淋巴组织中的表达水平最高。 dGK和1型单纯疱疹病毒胸苷激酶氨基酸序列的比对显示,dGK中还保留了五个区域,包括底物结合口袋和ATP结合甘氨酸环。据我们所知,这是克隆的线粒体核苷激酶的首次报道,也是两个哺乳动物脱氧核糖核苷激酶之间一般序列同源性的首次证明。我们的发现表明dCK和dGK在进化上相关,并且与疱疹病毒胸苷激酶家族有关。

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  • 作者

    Johansson, M; Karlsson, A;

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  • 年度 1996
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  • 正文语种 en
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