Heirwegh K P, Vermeir M, Molenberghs G
Laboratory of Hepatology, Catholic University of Leuven, Belgium.
J Biochem Biophys Methods. 1993 Sep;27(2):151-6. doi: 10.1016/0165-022x(93)90059-w.
Saturation curves for substrate interaction with an acceptor (enzyme, binding or carrier protein) are often analysed on the assumption that the amount of acceptor-bound substrate is negligible compared to its total amount. Analytical criteria permitting one to decide whether the assumption is justified or not for systems described by a single Michaelis-Menten equation have been derived previously. For more complicated systems, error formulae often cannot be obtained in closed form, and, if obtainable, are unwieldy. How this more complicated problem can be tackled is shown for mixtures of acceptor sites described by a sum of several Michaelis-Menten terms, without or with an additional term for 'non-specific' uptake or binding. In particular, it is shown that the maximum error, for which simple analytical expressions are obtained, provides a valid criterion for assessing whether substrate depletion is negligible or not.
底物与受体(酶、结合蛋白或载体蛋白)相互作用的饱和曲线分析通常基于这样的假设:与底物总量相比,受体结合的底物量可忽略不计。先前已推导了分析标准,可据此判断对于由单个米氏方程描述的系统,该假设是否合理。对于更复杂的系统,误差公式通常无法以封闭形式获得,即便能够获得,也很繁琐。本文针对由几个米氏项之和描述的受体位点混合物,展示了如何处理这个更复杂的问题,该混合物有无“非特异性”摄取或结合的附加项。特别指出的是,所获得的简单解析表达式的最大误差,为评估底物消耗是否可忽略不计提供了一个有效的标准。