Mimicking phosphorylation of Ser-74 on human deoxycytidine kinase selectively increases catalytic activity for dC and dC analogues
Mimicking phosphorylation of Ser-74 on human deoxycytidine kinase selectively increases catalytic activity for dC and dC analogues
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DOI：
10.1016/j.febslet.2008.01.048
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发表时间：
2008-03-05
期刊：
影响因子：
3.5
通讯作者：
Konrad, Manfred
中科院分区：
文献类型：
--
作者：
McSorley, Theresa;Ort, Stephan;Konrad, Manfred
dCK在Ser-74上的细胞内磷酸化导致核苷激酶活性增加。我们通过细菌产生的dCK中的Ser-74-Glu突变模拟了这种磷酸化，并使用各种核苷底物研究了动力学参数。S74E突变使dC的k（cat）值增加11倍，抗癌类似物dFdC和AraC的k（cat）值增加3倍。相反，嘌呤底物的速率降低。在REK293细胞中，通过比较瞬时转染的dCK(S74E)-GFP和野生型dCK-GFP，我们发现模拟Ser-74磷酸化对细胞定位没有影响。我们注意到磷酸化可能代表了一种机制来增强相对缓慢的dCK酶的催化活性。(c) 2008年欧洲生化学会联合会。Elsevier B.V.版权所有。
Intracellular phosphorylation of dCK on Ser-74 results in increased nucleoside kinase activity. We mimicked this phosphorylation by a Ser-74-Glu mutation in bacterially produced dCK and investigated kinetic parameters using various nucleoside substrates. The S74E mutation increases the k(cat) values 11-fold for dC, and 3-fold for the anti-cancer analogues dFdC and AraC. In contrast, the rate is decreased for the purine substrates. In REK293 cells, we found that by comparing transiently transfected dCK(S74E)-GFP and wild-type dCK-GFP, mimicking the phosphorylation of Ser-74 has no effect on cellular localisation. We note that phosphorylation may represent a mechanism to enhance the catalytic activity of the relatively slow dCK enzyme. (c) 2008 federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.