| 离子对反相HPLC测定人红细胞PRPP合成酶活性 |
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| 引用本文： | 彭岚,卢义钦,刘俊凡.离子对反相HPLC测定人红细胞PRPP合成酶活性[J].中国生物化学与分子生物学报,1997,13(1):113-115. |
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| 作者姓名： | 彭岚 卢义钦 刘俊凡 |
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| 作者单位： | 湖南医科大学血液生化研究室!长沙，410078，湖南医科大学血液生化研究室!长沙，410078，湖南医科大学血液生化研究室!长沙，410078 |
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| 基金项目： | 中华医学基金!会(CMB)资助900105 |
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| 摘 要： | 红细胞PRPP合成酶是红细胞内核苷酸代谢的关键酶，它参与嘌呤核苷酸的从头合成与补救合成途径，催化ATP与5-磷酸核糖（R5P）反应生成PRPP与AMP，而PRPP则是嘌呤嘧啶核苷酸合成途径的一个关键性中间产物．Stocchi等1]采用离子对反相高效液相色谱法（IPrHPLC）测定红细胞内ATP，ADP．AMP含量，曾获得满意的谱峰分离效果．Sakuma等2]应用rHPLC法测定了正常人及痛风等患者红细胞的PRPP合成酶活性，我们将Sakuma等的测酶技术与IPrHPLC法相结合，改用单液等度洗脱，达到了操作简便和灵敏、准确的要求．1材料和方法1．1试剂…
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| 收稿时间： | 1997-02-20 |
Estimation of Phosphoribosylpyrophosphate (PRPP) Synthetase Activity in Human Erythrocytes by Ion-pair rHPLC |
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| Peng Lan,Lu Yi-Qing,Liu Jun-Fan.Estimation of Phosphoribosylpyrophosphate (PRPP) Synthetase Activity in Human Erythrocytes by Ion-pair rHPLC[J].Chinese Journal of Biochemistry and Molecular Biology,1997,13(1):113-115. |
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| Authors: | Peng Lan Lu Yi-Qing Liu Jun-Fan |
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| Institution: | (Laboratory of Blood Biochemistry, Hunan Medical University, Changsha 410078 |
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| Abstract: | Using ion-pair reversed-phase HPLC (IPrHPLC), the PRPP synthetase activity inhuman erythrocytes was estimated. Methodological improvements have been taken on seeral aspects, including the removal of leukocytes and platelets in preparation of hemolysates, lysis of erythrocyte membranes and raising of efficiency in separation of ADP from ATP peaks etc. Thismethod fulfills the requirements of simplicity, reproducibility and accuracy. The intrabatch coefficient of variation was ±3. 0 % on average, the mean relative deviation was 2. 3 %, with a recoveryrate of 94.3—98.6%. The mean value of PRPP sythetase activities in the erythrocytes of 28healthy Chinese was 2. 19 ± 0. 61 μmol·min－1·g－1. |
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| Keywords: | 5-Phosphoribosyl-1-pyrophosphate (PRPP) synthetase Human erythrocytes Ionpair reversed-phase HPLC (IPrHPLC) |
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