Analysis of the interactions between an enzyme and multiple inhibitors using combination plots
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The concurrent effects of two enzyme inhibitors have been analysed previously with the Yonetani-Theorell plot to obtain insight into the interactions between bound inhibitors. This procedure, like many other traditional graphical methods in enzymology, is based on the estimation of intersecting tendencies in a family of lines. In a recent paper from this laboratory [Chan (1995) Biochem. J. 311, 981-985] it was shown that a plot of this nature may sometimes be replaced, with advantage, by a 'combination plot' in which all data points are accommodated in a single line. We have now extended this approach to analyse the effects of multiple inhibitors and have developed combination plots which illustrate the interaction behaviour in an optimal manner. Thus, in these plots, the synergistic or antagonistic nature of the interactions is clearly evident from the slope, which also provides a direct estimate of the interaction coefficient. The analysis is more efficient and consequently requires fewer enzyme assays. This approach is applicable to various special cases, including that in which three inhibitors bind simultaneously to the enzyme.
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