Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis
文献类型：期刊论文
| 作者 | Wang, Tongdan1,2; Kang, Jingwu1 |
| 刊名 | Electrophoresis
 |
| 出版日期 | 2009-04-01 |
| 卷号 | 30期号:8页码:1349-1354 |
| 关键词 | Ce Electrophoretically mediated microanalysis Hexokinase Kinase inhibitor screening |
| ISSN号 | 0173-0835 |
| DOI | 10.1002/elps.200800542 |
| 通讯作者 | Kang, jingwu(jingwu.kang@mail.sioc.ac.cn) |
| 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 mgcl(2), 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. |
| WOS关键词 | MICELLAR ELECTROKINETIC CHROMATOGRAPHY ; FLOW CAPILLARY-ELECTROPHORESIS ; PARTIAL FILLING TECHNIQUE ; ENANTIOMERIC SEPARATION ; ENZYMATIC-REACTION ; NATURAL EXTRACTS ; VANCOMYCIN ; ASSAYS ; VALIDATION ; CANCERS |
| WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry |
| WOS类目 | Biochemical Research Methods ; Chemistry, Analytical |
| 语种 | 英语 |
| WOS记录号 | WOS:000265968300009 |
| 出版者 | WILEY-V C H VERLAG GMBH |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2404256 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Kang, Jingwu |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Organ Chem, Shanghai 200032, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Tongdan,Kang, Jingwu. Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis[J]. Electrophoresis,2009,30(8):1349-1354. |
| APA | Wang, Tongdan,&Kang, Jingwu.(2009).Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis.Electrophoresis,30(8),1349-1354. |
| MLA | Wang, Tongdan,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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来源：中国科学院大学
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