Development of a capillary electrophoresis method for analyzing adenosine deaminase and purine nucleoside phosphorylase and its application in inhibitor screening

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• 作者：Yanfei Qi ; Youxin Li ; ^{lyx@tju.edu.cn" class="auth_mail" title="E-mail the corresponding author} ; ^{liyouxin2008@163.com" class="auth_mail" title="E-mail the corresponding author} ; James J. Bao
• 关键词：Capillary electrophoresis ; Adenosine deaminase ; Purine nucleoside phosphorylase ; Inhibitor screening ; Red blood cells ; Traditional Chinese medicines
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：1 August 2016
• 年：2016
• 卷：506
• 期：Complete
• 页码：31-44
• 全文大小：960 K

文摘

A novel capillary electrophoresis (CE) method was developed for simultaneous analysis of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) in red blood cells (RBCs). The developed method considered and took advantage of the natural conversion from the ADA product, inosine to hypoxanthine. The transformation ratio was introduced for ADA and PNP analysis to obtain more reliable results. After optimizing the enzymatic incubation and electrophoresis separation conditions, the determined activities of ADA and PNP in 12 human RBCs were 0.237–0.833 U/ml and 9.013–10.453 U/ml packed cells, respectively. The analysis of ADA in mice RBCs indicated that there was an apparent activity difference between healthy and hepatoma mice. In addition, the proposed method was also successfully applied in the inhibitor screening from nine traditional Chinese medicines, and data showed that ADA activities were strongly inhibited by Rhizoma Chuanxiong and Angelica sinensis. The inhibition effect of Angelica sinensis on ADA is first reported here and could also inhibit PNP activity.