Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis

CORC > 上海有机化学研究所 > 中国科学院上海有机化学研究所 > 分析化学研究室

	Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis
	Wang TD(汪彤丹) ; Kang JW(康经武)
刊名	Electrophoresis
	2009
卷号	30 期号:8 页码:1349-1354
ISSN号	0173-0835
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
通讯作者	康经武
英文摘要	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.
学科主题	分析化学
收录类别	SCI
原文出处	http://dx.doi.org/10.1002/elps.200800542
语种	英语
WOS记录号	WOS:000265968300009
公开日期	2013-08-29
内容类型	期刊论文
源URL	[http://202.127.28.38/handle/331003/28685]
专题	上海有机化学研究所_分析化学研究室
推荐引用方式 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.