The 'enzyme-probe' method for characterizing metabolite pools. The use of NAD-glycohydrolase in human erythrocyte sonicate as a model system.
The 'enzyme-probe' method for characterizing metabolite pools. The use of NAD-glycohydrolase in human erythrocyte sonicate as a model system.
复制标题
用于表征代谢物池的“酶探针”方法。
DOI：
10.1111/j.1432-1033.1979.tb12996.x
复制
发表时间：
1979
期刊：
影响因子：
--
通讯作者：
P. Friedrich
中科院分区：
文献类型：
--
作者：
M. Solti;P. Friedrich
1.描述了用于测试代谢物池的均匀性的方法。在所研究的体系中引入了一种可以攻击代谢产物的外源酶。通过分析代谢物分解的时间过程，可以决定合并物在对探针-酶的反应性方面是否均匀，或者是否可以分为不同反应性的部分。2.以NAD-糖水解酶为探针，以人红细胞超声波破碎物为模型系统，说明了从这些实验中获得的信息。在浓缩的超声处理液中的烟酰胺腺嘌呤二核苷酸池可以被分解成三个部分（I，II和III），半衰期分别约为1，7和240分钟。组分I是游离NAD，组分II是与甘油醛-3-磷酸脱氢酶结合的NAD，组分III是与一些迄今未鉴定的蛋白质强烈结合的辅酶。超声糖酵解似乎只需要馏分II，并且在所应用的实验条件下不能使用馏分III。3.讨论了酶探针法的适用范围。
1. An approach for testing the homogeneity of metabolite pools is described. An alien enzyme that can attack the metabolite in question is introducted into the system studied. By analyzing the time-course of decomposition of the metabolite it can be decided whether the pool is homogeneous in respect of reactivity towards the probe-enzyme or can be divided into fractions of different reactivities. 2. The information obtainable from such experiments is illustrated by the case of human erythrocyte sonicate as model system with NAD-glycohydrolase as probe-enzyme. The nicotinamide adenine dinucleotide pool in the concentrated sonicate could be resolved into three fractions (I, II and III) with half-lives of about 1, 7 and 240 min, respectively. Fraction I is free NAD, fraction II is NAD bound to glyceraldehyde-3-phosphate dehydrogenase, and fraction III is coenzyme strongly bound to some, so far unidentified, protein. Sonicate glycolysis seems to require only fraction II and is unable to use fraction III under the experimental conditions applied. 3. The scope of application of the enzyme-probe method is discussed.