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Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis
其他题名基于二磷酸腺苷(ADP)检测的电泳介导微分析技术用于己糖激酶抑制剂的筛选
Wang TD(汪彤丹); Kang JW(康经武)
2009
发表期刊Electrophoresis
ISSN0173-0835
卷号30期号:8页码:1349-1354
摘要A CE-based method for hexokinase inhibitor screening was developed in the present paper, in this method, hexokinase activity was assayed via electrophoretically mediated microanalysis (EMMA), which combines on-column hexokinase-mediated reaction and measurement of produced adenosine 5'-diphosphate (ADP) via electrophoretical separation and UV detection. Enzyme inhibition can be read out directly from the reduced peak area of ADP in comparison with a reference electropherogram obtained in the absence of any inhibitor. Conditions for on-column enzyme reaction and separation of adenosine 5'-triphosphate (ATP) and ADP were optimized. The optimal buffer composition for enzymatic reaction was 25 mM HEPFS buffer (pH 7.5) containing 5 mM MgCl2, whereas the optimal buffer composition for separation was 100 in M Tris-phosphate buffer (pH 5.5) containing 0.02% (m/v) hexadimethrine bromide (HDB). Fortunately, discontinuous buffer system call be adapted easily in the EMMA method. The time for separation was reduced dramatically to less than 3 min by reversing the direction of EOF via dynamically coating the capillary wall with the cationic polyelectrolyte HDB. Moreover, the peak tailing of ATP was also reduced by HDB coating. The Z' factor as high as 0.98 was obtained, indicating a high quality of the screening data. The present method is simple, robust and cost-effective.
学科领域分析化学
部门归属上海有机所; 中科院研究生院
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收录类别SCI
语种英语
WOS记录号WOS:000265968300009
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被引频次:22[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.sioc.ac.cn/handle/331003/28685
专题分析化学研究室
通讯作者Kang JW(康经武)
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GB/T 7714
Wang TD,Kang JW. Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis[J]. Electrophoresis,2009,30(8):1349-1354.
APA 汪彤丹,&康经武.(2009).Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis.Electrophoresis,30(8),1349-1354.
MLA 汪彤丹,et al."Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis".Electrophoresis 30.8(2009):1349-1354.
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