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
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
070307 [化学生物学];
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
相关论文
共 20 条
[1]
Agarwal K C, 1978, Methods Enzymol, V51, P483
[2]
MAMMALIAN DEOXYRIBONUCLEOSIDE KINASES [J].
ARNER, ESJ ;
ERIKSSON, S .
PHARMACOLOGY & THERAPEUTICS, 1995, 67 (02) :155-186
[3]
DIFFERENT REGULATION OF THE HUMAN THYMIDINE KINASE PROMOTER IN NORMAL HUMAN-DIPLOID IMR-90 FIBROBLASTS AND HELA-CELLS [J].
CHANG, ZF ;
HUANG, DY ;
LAI, TC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (45) :27374-27379
[4]
Exploring acyclic nucleoside analogues as inhibitors of Mycobacterium tuberculosis thymidylate kinase [J].
Familiar, Olga ;
Munier-Lehmann, Helene ;
Negri, Ana ;
Gago, Federico ;
Douguet, Dominique ;
Rigouts, Leen ;
Hernandez, Ana-Isabel ;
Camarasa, Maria-Jose ;
Perez-Perez, Maria-Jesus .
CHEMMEDCHEM, 2008, 3 (07) :1083-1093
[5]
Substituted benzyl-pyrimidines targeting thymidine monophosphate kinase of Mycobacterium tuberculosis:: Synthesis and in vitro anti-mycobacterial activity [J].
Gasse, Cecile ;
Douguet, Dominique ;
Huteau, Valerie ;
Marchal, Gilles ;
Munier-Lehmann, Helene ;
Pochet, Sylvie .
BIOORGANIC & MEDICINAL CHEMISTRY, 2008, 16 (11) :6075-6085
[6]
Role of antimetabolites of purine and pyrimidine nucleotide metabolism in tumor cell differentiation [J].
Hatse, S ;
De Clercq, E ;
Balzarini, J .
BIOCHEMICAL PHARMACOLOGY, 1999, 58 (04) :539-555
[7]
Synthetic lethality by lentiviral short hairpin RNA silencing of thymidylate kinase and doxorubicin in colon cancer cells regardless of the p53 status [J].
Hu, Chun-Mei ;
Chang, Zee-Fen .
CANCER RESEARCH, 2008, 68 (08) :2831-2840
[8]
HUMAN DTMP KINASE - GENE-EXPRESSION AND ENZYMATIC-ACTIVITY COINCIDING WITH CELL-CYCLE PROGRESSION AND CELL-GROWTH [J].
HUANG, SH ;
TANG, A ;
DRISCO, B ;
ZHANG, SQ ;
SEEGER, R ;
LI, C ;
JONG, A .
DNA AND CELL BIOLOGY, 1994, 13 (05) :461-471
[9]
[10]
LEE LS, 1977, J BIOL CHEM, V252, P5686