A simple and rapid method for measuring alpha-D-phosphohexomutases activity by using anion-exchange chromatography coupled with an electrochemical detector
文献类型：期刊论文
| 作者 | Jia, Xiaochen1,2; Kang, Jian3; Yin, Heng1 |
| 刊名 | PEERJ
 |
| 出版日期 | 2016-01-05 |
| 卷号 | 4 |
| 关键词 | Hpaec-pad N-acetylglucosamine-phosphate Mutase N-acetylglucosamine-1-phosphate N-acetylglucosamine-6-phosphate Alpha-d-phosphohexomutases |
| ISSN号 | 2167-8359 |
| DOI | 10.7717/peerj.1517 |
| 文献子类 | Article |
| 英文摘要 | The interconversion of hexose-6-phosphate and hexose-l-phosphate can be directly analyzed by high-performance anion-exchange chromatography coupled with an electrochemical detector (HPAEC-PAD). Thus, this method can be used to measure the activities of N-acetylglucosamine-phosphate mutase (AGM), glucosamine-phosphate mutase (GlmM) and phosphoglucornutase (PGM), which are the members of alpha-D-phosphohexomutases superfamily. The detection limits were extremely low as 2.747 pmol, 1.365 pmol, 0.512 pmol, 0.415 pmol, 1.486 pmol and 0.868 pmol for N-acetylglucosamine-1-phosphate (GleNAc-1-P), N-acetylglucosamine-6-phosphate (GkNAc-6-P), glucosamine-1-phosphate ( GlcN-1 -P), glucosamine-6-phosphate (GkN-6-P), glucose-1-phosphate (Glc-1-P) and glucose-6-phosphate (Gk-6-P), respectively. By employing HPAEC-PAD, activities of AtAGM (AGM from Arabidopsis thaliana) on these six phosphohexoses can be detected. The K-m of AtAGM on Glc-1-P determined by HPAEC-PAD was 679.18 +/- 156.40 mu M, which is cornparable with the K-m of 707.09 +/- 170.36 mu M detected by traditional coupled assay. Moreover, the activity of MtGlmM (G1mM from Mycobacterium tuberculosis) on GlcN-6-P tested by HPAEC-PAD was 7493.40 +/- 309.12 nmol/min.mg, which is much higher than 288.97 +/- 35.28 nmol/min.mg obtained by the traditional coupled assay. Accordingly, HPAEC-PAD is a more rapid and simple method than the traditional coupled assays given its high specificity and sensitivity, and will certainly bring convenience to further research of alpha-D-phosphohexomutases. |
| WOS关键词 | ESCHERICHIA-COLI ; O-GLCNAC ; N-ACETYLGLUCOSAMINE-1-PHOSPHATE URIDYLTRANSFERASE ; PHOSPHOGLUCOSAMINE MUTASE ; SUGAR PHOSPHATES ; PATHWAY ; GLMU ; GLYCOSYLATION ; STRESS ; YEAST |
| WOS研究方向 | Science & Technology - Other Topics |
| 语种 | 英语 |
| WOS记录号 | WOS:000368065500003 |
| 出版者 | PEERJ INC |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/171506]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 通讯作者 | Yin, Heng |
| 作者单位 | 1.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.Dalian Med Univ, Dept Biochem & Mol Biol, Dalian, Peoples R China |
| 推荐引用方式 GB/T 7714 | Jia, Xiaochen,Kang, Jian,Yin, Heng. A simple and rapid method for measuring alpha-D-phosphohexomutases activity by using anion-exchange chromatography coupled with an electrochemical detector[J]. PEERJ,2016,4. |
| APA | Jia, Xiaochen,Kang, Jian,&Yin, Heng.(2016).A simple and rapid method for measuring alpha-D-phosphohexomutases activity by using anion-exchange chromatography coupled with an electrochemical detector.PEERJ,4. |
| MLA | Jia, Xiaochen,et al."A simple and rapid method for measuring alpha-D-phosphohexomutases activity by using anion-exchange chromatography coupled with an electrochemical detector".PEERJ 4(2016). |
入库方式： OAI收割
来源：大连化学物理研究所
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