Roberts M W, Preiss J, Okita T W
Institute of Biological Chemistry, Washington State University, Pullman 99163-6340, USA.
Anal Biochem. 1995 Feb 10;225(1):121-6. doi: 10.1006/abio.1995.1117.
Conditions are described for a capillary zone electrophoresis-based assay of ADP-glucose pyrophosphorylase, the first enzyme in the de novo synthesis for alpha-glucan synthesis in plants and bacteria. ATP, ADP-glucose, and other adenine derivatives are effectively resolved and quantified in 12 min of electrophoresis. The method requires minimal sample manipulations and no postincubation hydrolyses, is effective over a wide range of enzyme and substrate concentrations, has a sensitivity comparable to radioisotope assays, and yields reproducible results in either the forward and reverse directions. Moreover, the assay is applicable to other enzymes that utilize nucleosides or their bases as substrates or products.
本文描述了一种基于毛细管区带电泳的ADP-葡萄糖焦磷酸化酶检测方法，该酶是植物和细菌中α-葡聚糖从头合成的首个酶。在12分钟的电泳过程中，ATP、ADP-葡萄糖和其他腺嘌呤衍生物能够得到有效分离和定量。该方法所需的样品处理最少，无需孵育后水解，在广泛的酶和底物浓度范围内均有效，灵敏度与放射性同位素检测相当，并且正反两个方向均可产生可重复的结果。此外，该检测方法适用于其他利用核苷或其碱基作为底物或产物的酶。
