RAPID MEASUREMENT OF BINDING CONSTANTS AND HEATS OF BINDING USING A NEW TITRATION CALORIMETER
RAPID MEASUREMENT OF BINDING CONSTANTS AND HEATS OF BINDING USING A NEW TITRATION CALORIMETER
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DOI：
10.1016/0003-2697(89)90213-3
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发表时间：
1989-05-15
影响因子：
2.9
通讯作者：
LIN, LN
中科院分区：
文献类型：
--
作者：
WISEMAN, T;WILLISTON, S;LIN, LN
本文介绍了一种新的滴定量热计，并给出了2“-单磷酸胞苷（2”CMP）与核糖核酸酶A活性位点结合的结果。该仪器的特点包括非常高的灵敏度，快速的量热响应和快速的热平衡。方便的软件可用于仪器操作、数据收集、数据简化和去卷积以获得结合参数n、Δ的最小二乘估计。H °，Δ S °，以及结合常数K。该仪器的样品吞吐量高，在有利的条件下，结合常数可高达108 M-1。研究了牛核糖核酸酶A（RNase）/2“CMP系统在50倍RNase浓度范围内和两种不同温度下的反应。结合常数在105至106 M-1的范围内，取决于条件，结合热约为100。-15，000卡/摩尔。重复测定表明n，Δ H °，和K值在最有利的浓度范围内。
A new titration calorimeter is described and results are presented for the binding of cytidine 2''-monophosphate (2''CMP) to the active site of ribonuclease A. The instrument characteristics include very high sensitivity, rapid calorimetric response, and fast thermal equilibration. Convenient software is available for instrument operation, data collection, data reduction, and deconvolution to obtain least-squares estimates of binding parameters n, .DELTA. H.degree., .DELTA.S.degree., and the binding constant K. Sample through-put for the instrument is high, and under favorable conditions binding constants as large as 108 M-1 can be measured. The bovine ribonuclease A (RNase)/2''CMP system was studied over a 50-fold range of RNase concentration and at two different temperatures. The binding constants were in the 105 to 106 M-1 range, depending on conditions, and heats of binding ca. -15,000 cal/mol. Repeat determinations suggested errors of only a few percent in n, .DELTA.H.degree., and K values over the most favorable concentration range.