代谢途径的动力学。一种用于研究通量控制的体外系统。
Kinetics of metabolic pathways. A system in vitro to study the control of flux.
作者信息
Torres N V, Mateo F, Meléndez-Hevia E, Kacser H
出版信息
Biochem J. 1986 Feb 15;234(1):169-74. doi: 10.1042/bj2340169.
Abstract
A method for determining Control Coefficients is proposed for systems studied in vitro and applied to a model pathway. Rat liver extract, which converts glucose into glycerol 3-phosphate, was used with the addition to the incubation mixture of fructose-bisphosphate aldolase, triose-phosphate isomerase and glycerol-3-phosphate dehydrogenase as 'auxiliary' enzymes, which leaves all the control on the first three enzymes. The flux of the metabolic pathway was recorded by assaying NADH decay. Flux Control Coefficients (CJE) of hexokinase, glucose-6-phosphate isomerase and phosphofructokinase were calculated by titration of the system with increasing quantities of extraneous enzymes. It is shown that the summation property is fulfilled. The applicability of this procedure to study the control in any metabolic pathway is discussed. Possible relevance of the method to conditions in vivo and its limitations are considered.
摘要
本文提出了一种用于体外研究系统的控制系数测定方法，并将其应用于一个模型途径。将能把葡萄糖转化为3-磷酸甘油的大鼠肝脏提取物与果糖二磷酸醛缩酶、磷酸丙糖异构酶和3-磷酸甘油脱氢酶作为“辅助”酶添加到孵育混合物中，这样所有的控制都集中在前三种酶上。通过检测NADH的衰减来记录代谢途径的通量。通过用不断增加量的外源酶滴定系统来计算己糖激酶、葡萄糖-6-磷酸异构酶和磷酸果糖激酶的通量控制系数（CJE）。结果表明该方法满足加和性。讨论了该程序在研究任何代谢途径控制方面的适用性。考虑了该方法与体内条件的可能相关性及其局限性。
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