THEORETICAL ASPECTS OF SPECIFIC AND NONSPECIFIC EQUILIBRIUM BINDING OF PROTEINS TO DNA AS STUDIED BY THE NITROCELLULOSE FILTER BINDING ASSAY - COOPERATIVE AND NON-COOPERATIVE BINDING TO A ONE-DIMENSIONAL LATTICE
THEORETICAL ASPECTS OF SPECIFIC AND NONSPECIFIC EQUILIBRIUM BINDING OF PROTEINS TO DNA AS STUDIED BY THE NITROCELLULOSE FILTER BINDING ASSAY - COOPERATIVE AND NON-COOPERATIVE BINDING TO A ONE-DIMENSIONAL LATTICE
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DOI：
10.1016/0022-2836(82)90481-8
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发表时间：
1982-01-01
影响因子：
5.6
通讯作者：
DAVIES, RW
中科院分区：
文献类型：
--
作者：
CLORE, GM;GRONENBORN, AM;DAVIES, RW
平衡结合等温线的分析方法，如测定分数的硝化纤维素过滤器结合试验，得到。对于至少有1个蛋白质分子结合的DNA，作为游离蛋白浓度（LF）的函数，需要一种不同于分析传统平衡结合数据所需的理论框架，在传统平衡结合数据中，结合蛋白质的摩尔数/mol DNA， theta.c，作为游离蛋白浓度的函数来测量。分析由测量。θ产生的平衡绑定数据所需的理论框架。（LF）是为蛋白质与大量非特异性位点的合作和非合作结合以及在DNA分子上存在大量非特异性位点的情况下与特定位点的合作和非合作结合而开发的。理论应用起来很简单，方程。（LF）易于推导和求值，适用于最小二乘分析。本文给出了EcoRI限制性内切酶与噬菌体的特异性和非特异性结合的两个应用该理论分析实验数据的例子。对于干酪乳杆菌二氢叶酸还原酶与pBR322和pWDLcB1 DNA的特异性和非特异性结合，后者与前者仅差异2.9倍。103碱基对插入含有干酪乳杆菌二氢叶酸还原酶结构基因。测量θ的理论和实验优缺点。（LF）而不是。theta。c （LF）。
The analysis of equilibrium binding isotherms obtained by methods such as the nitrocellulose filter binding assay, which measure the fraction, .theta., of DNA to which at least 1 protein molecule is bound, as a function of the free protein concentration (LF) require a different type of theoretical framework from that required for analysis of conventional equilibrium binding data, in which the number of moles of protein bound/mol of DNA, .theta.c, is measured as a function of LF. The theoretical framework required to analyse equilibrium binding data generated by measuring .theta.(LF) is developed for cooperative and noncooperative binding of a protein to a large number of non-specific sites and to a specific site(s) in the presence of a large number of non-specific sites on a DNA molecule. The theory is simple to apply, equations for .theta.(LF) being easy to derive and evaluate, and is suitable for least-squares analysis. Two examples of the application of the theory to the analysis of experimental data are provided for the specific and non-specific binding of the EcoRI restriction endonuclease to bacteriophage .lambda. DNA, and for the specific and non-specific binding of the enzyme dihydrofolate reductase from Lactobacillus casei to pBR322 and pWDLcB1 DNA, the latter differing from the former only in a 2.9 .times. 103 base-pair insert containing the L. casei dihydrofolate reductase structural gene. The theoretical and experimental advantages and disadvantages of measuring .theta.(LF) rather than .theta.C(LF) are discussed.