A bioluminescent method for measuring thymidylate kinase activity suitable for high-throughput screening of inhibitor

被引：6
|
作者：
Hu, Chun-Mei [1 ]
Chang, Zee-Fen [1 ]
机构：
[1] Natl Taiwan Univ, Coll Med, Inst Biochem & Mol Biol, Taipei 10617, Taiwan
来源：
ANALYTICAL BIOCHEMISTRY | 2010年 / 398卷 / 02期
关键词：
THYMIDINE MONOPHOSPHATE KINASE; MYCOBACTERIUM-TUBERCULOSIS; NUCLEOSIDE ANALOGS; DNA-SYNTHESIS; GENE; DESIGN; PURINE; GROWTH; CELLS;
D O I：
10.1016/j.ab.2009.12.001
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Blocking human thymidylate kinase (TMPK) function has a chemosensitization effect in anticancer treatment. However, a rapid and sensitive TMPK activity assay method suitable for inhibitor screening has been lacking. We have designed a luciferase-coupled TMPK assay in which luminescence emission is proportional to the magnitude of TMPK inhibition. The advantages of using this new method over the conventional nicotinamide adenine dinucleotide (reduced form, NADH)-coupling method in screening inhibitor include low cost, low limit in detecting inhibitory signal, more accurate, and devoid of interference due to compound absorbance at 340 nm. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码：269 / 271
页数：3
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