Bessho M, Miyauchi Y, Sano N, Nakamura T, Tajima T
Eisai Research Laboratories, Eisai Co., Ltd., Tokyo, Japan.
J Nutr Sci Vitaminol (Tokyo). 1988 Dec;34(6):607-14. doi: 10.3177/jnsv.34.607.
An assay method for pyruvate kinase in rat plasma is described. Plasma samples were incubated with ADP and phosphoenolpyruvate in Tris buffer solution. The ATP produced by pyruvate kinase was measured by photocounting after the addition of a commercially available luciferin-luciferase preparation. Interference by ATP or adenylate kinase originally present in the sample was removed by a high degree of dilution. The assay is sensitive, reproducible, and rapid, especially when used for large numbers of samples. By this method, pyruvate kinase activity in normal rats was determined to be 0.51 +/- 0.05 (n = 6) U/ml plasma. In rats fed a vitamin E-deficient basal diet for 7, 10, or 14 weeks, pyruvate kinase activities were 0.70 +/- 0.11, 1.64 +/- 0.51, and 4.28 +/- 0.85 (n = 6) U/ml plasma, respectively. This method appears to be useful for the determination of pyruvate kinase activity in nutritional or pharmacological studies.
本文描述了一种大鼠血浆中丙酮酸激酶的检测方法。血浆样本在Tris缓冲溶液中与ADP和磷酸烯醇丙酮酸一起孵育。在加入市售的荧光素-荧光素酶制剂后，通过光计数法测定丙酮酸激酶产生的ATP。通过高度稀释去除样品中原本存在的ATP或腺苷酸激酶的干扰。该检测方法灵敏、可重复且快速，尤其适用于大量样本。通过这种方法，正常大鼠血浆中丙酮酸激酶活性测定为0.51±0.05（n = 6）U/ml。在喂食维生素E缺乏基础饮食7周、10周或14周的大鼠中，丙酮酸激酶活性分别为0.70±0.11、1.64±0.51和4.28±0.85（n = 6）U/ml血浆。该方法似乎可用于营养或药理学研究中丙酮酸激酶活性的测定。
