线粒体和胞质胸苷激酶（TK2和TK1）对5-溴乙烯基2'-脱氧尿苷的磷酸化作用及其在粗提物中TK2活性选择性测定中的应用。
5-Bromovinyl 2'-deoxyuridine phosphorylation by mitochondrial and cytosolic thymidine kinase (TK2 and TK1) and its use in selective measurement of TK2 activity in crude extracts.
作者信息
Wang Liya, Eriksson Staffan
机构信息
Department of Anatomy, Physiology and Biochemistry, Section of Veterinary Medical Biochemistry, Swedish University of Agricultural Sciences, The Biomedical Centre, Uppsala, Sweden.
出版信息
Nucleosides Nucleotides Nucleic Acids. 2008 Jun;27(6):858-62. doi: 10.1080/15257770802146510.
Mitochondrial thymidine kinase (TK2) is responsible for phosphorylation of thymidine and deoxycytidine and plays a crucial role in mitochondrial DNA precursor synthesis. TK2 is expressed in all tissues at low levels complicating accurate determinations, especially in tissues with high cytosolic thymidine kinase (TK1) activity. Recently, 5-bromovinyl 2 '-deoxyuridine (BvdU) at 0.2 micro M was used to measure TK2 activity selectively. BvdU phosphorylation by pure human TK2 and TK1 was tested here, and the ratio of BvdU phosphorylation by TK2/TK1 was 91 at 0.2 micro M but was 500 at 2.5 micro M. Therefore, for reliable measurement of TK2 activity higher BvdU concentration should be used.
线粒体胸苷激酶（TK2）负责胸苷和脱氧胞苷的磷酸化，在线粒体DNA前体合成中起关键作用。TK2在所有组织中低水平表达，这使得准确测定变得复杂，尤其是在具有高胞质胸苷激酶（TK1）活性的组织中。最近，使用0.2微摩尔的5-溴乙烯基2'-脱氧尿苷（BvdU）来选择性测量TK2活性。在此测试了纯人TK2和TK1对BvdU的磷酸化，在0.2微摩尔时TK2/TK1对BvdU的磷酸化比率为91，但在2.5微摩尔时为500。因此，为了可靠地测量TK2活性，应使用更高浓度的BvdU。
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