Computer programs for the rapid determination of enzyme kinetics on MS-DOS compatible microcomputers.
Computer programs for the rapid determination of enzyme kinetics on MS-DOS compatible microcomputers.
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用于在 MS-DOS 兼容微型计算机上快速测定酶动力学的计算机程序。
DOI：
10.1093/bioinformatics/6.1.51
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Maloy,S
中科院分区：
文献类型：
--
作者：
Myers,R;Olson,S;Maloy,S
分子生物学家和生物化学家定期想要确定酶的活性和动力学。为了确定活性，通常用手绘制数据图并用眼睛拟合。为了确定酶促反应的动力学，对活性数据进行各种重新绘图（也是手绘的）并通过目测拟合以估计动力学参数。这些研究酶的图形方法是艰巨的，容易出现线拟合错误。专业的酶动力学家（如今有些罕见）很少绘制曲线图，而是依靠强大的计算机程序，将数据的数值分析与各种统计程序结合起来。这些程序通常在小型计算机和分时机器上运行，这对于通常不需要大量计算能力的研究人员来说并不方便。相比之下，大多数实验室都有微型计算机，分子生物学家越来越习惯于用它们进行文字处理和数据分析。为此，我们在MS-DOS兼容微机上编写了两个FORTRAN程序，用于快速计算酶动力学参数。为了确定酶对给定底物的特异性（Vi）、最大速度（Kmax）和亲和力（Km），从几种底物浓度下酶反应的初始速度（Vi）计算动力学参数（5，）（Cornish-Bowden，1979）。程序MEDIAN和HYPER是先前公布的程序的变体（Cornish-Bowden和Eisenthal，1974; Cornish-Bowden等人，Cleland，1963）用于从这些数据确定酶动力学。我们用IBM/Ryan-McFarland Professional FORTRAN语言在FORTRAN 77中重写了它们，这样它们就可以在现在普遍存在的MS-DOS兼容的微型计算机上运行。由于我们使用了这个特殊的FORTRAN编译器，所以需要一个数学协处理器来运行程序。程序HYPER通过酶活性数据与Michaelis-Menten方程定义的双曲线的最小二乘拟合来计算Km、Vmix及其标准误差：
Molecular biologists and biochemists periodically want to determine the activity and kinetics of enzymes. In order to determine the activity, plots of the data are usually drawn by hand and fitted by eye. In order to determine the kinetics of the enzymatic reaction, the activity data are subjected to various replots (also hand-drawn) and fitted by eye to estimate the kinetic parameters. These graphical approaches to study enzymes are arduous and prone to line-fitting errors. Professional enzyme kineticists (somewhat rare these days) infrequently draw plots, relying instead on powerful computer programs that combine numerical analysis of the data with various statistical routines. These programs often run on minicomputers and time-sharing machines which are not convenient for researchers that do not usually require large amounts of computation power. In contrast, microcomputers are found in most laboratories, and molecular biologists are becoming increasingly comfortable with them for word processing and data analysis. With this in mind, we wrote two FORTRAN progams on MS-DOS compatible microcomputers for quickly calculating enzyme kinetic parameters. In order to determine the specificity (V^ JK^, maximum velocity (Kmax), and affinity (Km) of an enzyme for a given substrate, the kinetic parameters are calculated from the initial velocity (V,) of the enzymatic reaction at several substrate concentrations (5,)(Cornish-Bowden, 1979). The programs MEDIAN and HYPER are variations of previously published programs (Cornish-Bowden and Eisenthal, 1974; Cornish-Bowden et al., Cleland, 1963) for determining enzyme kinetics from such data. We rewrote them in FORTRAN 77 using IBM/Ryan-McFarland Professional FORTRAN so they could be run on the now ubiquitous MS-DOS compatible microcomputers. Since we used this particular FORTRAN compiler, a math co-processor is required to run the programs. The program HYPER calculates Km, VmiX and their standard errors by a least-squares fit of enzyme activity data to a hyperbola defined by the Michaelis-Menten equation: