title: Development of a capillary electrophoresis method for analyzing adenosine deaminase and purine nucleoside phosphorylase and its application in inhibitor screening.
abstract: 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.
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery and High Efficiency, Collaborative Innovation Center of Chemical Science and Engineering, School of Pharmaceutical Science and Technology, Tianjin University, Nankai, Tianjin 300072, China.
authors.forename: Yanfei
authors.initials: Y
authors.lastname: Qi
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery and High Efficiency, Collaborative Innovation Center of Chemical Science and Engineering, School of Pharmaceutical Science and Technology, Tianjin University, Nankai, Tianjin 300072, China. Electronic address: lyx@tju.edu.cn.
authors.forename: Youxin
authors.initials: Y
authors.lastname: Li
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery and High Efficiency, Collaborative Innovation Center of Chemical Science and Engineering, School of Pharmaceutical Science and Technology, Tianjin University, Nankai, Tianjin 300072, China.
authors.forename: James J
authors.initials: JJ
authors.lastname: Bao
mesh_terms: D000243:Adenosine Deaminase / Q000032:analysis* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D019075:Electrophoresis, Capillary / Q000379:methods*; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000494:pharmacology*; D006801:Humans; D008516:Medicine, Chinese Traditional; D011683:Purine-Nucleoside Phosphorylase / Q000032:analysis* / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D004791:Enzyme Inhibitors; D011683:Purine-Nucleoside Phosphorylase; D000243:Adenosine Deaminase
keywords: Adenosine deaminase; Capillary electrophoresis; Inhibitor screening; Purine nucleoside phosphorylase; Red blood cells; Traditional Chinese medicines