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双倒数图

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Lineweaver-Burke plot.svg

双倒数图也称为莱恩威弗-伯克作图,是生物化学用来描述酶动力学的莱恩威弗-伯克方程的图示法,由汉斯·莱恩威弗英语Hans Lineweaver迪恩·伯克英语Dean Burk于1934年提出[1]

推导

双倒数图被用来图像分析米氏方程

取倒数得到

其中

V反应速率
Km为米氏常数。
Vmax为最大反应速率。
[S]为底物浓度

应用

在强大的计算机和非线性回归软件出现前双倒数图被广泛用来确定酶动力学里的项,比如KmVmax。双倒数图的截距等于Vmax的逆数。双倒数图的等于−1/Km。双倒数图还能很快地体现不同形式的酶抑制。

双倒数图扭曲数据结构,因此它不能可靠地确定酶动力学系数。虽然今天它依然被用来显示动力学数据[2],一般米氏动力学的非线性回归图象或者其它线性图象如哈尼斯-伍尔夫图英语Hanes–Woolf plot伊迪-霍夫斯蒂图英语Eadie–Hofstee diagram被用来计算系数[3]

双倒数图可以用来区别竞争性抑制非竞争性抑制英语Non-competitive inhibition不竞争性抑制英语Uncompetitive inhibitor。竞争性抑制剂和不竞争性抑制剂的y截距相同,但是倾斜度不同,x也不同。非竞争性抑制剂和不竞争性抑制剂的x截距相同,但是倾斜度不同,因此y不同。竞争性抑制剂和非竞争性抑制剂的yx都不同。

缺点

在较旧的书籍里双倒数图经常被使用,但是它很容易出错误。它的y轴是反应速度的倒数,因此小的测量错误会被放大。此外大多数图在y轴的右边很远的地方,因此要通过很大的外推来获得xy的截距[4]

参考资料

  1. ^ Lineweaver, H and Burk, D. The Determination of Enzyme Dissociation Constants. Journal of the American Chemical Society. 1934, 56 (3): 658–666. doi:10.1021/ja01318a036. 
  2. ^ Hayakawa, K.; Guo, L.; Terentyeva, E.A.; Li, X.K.; Kimura, H.; Hirano, M.; Yoshikawa, K.; Nagamine, T.; et al. Determination of specific activities and kinetic constants of biotinidase and lipoamidase in LEW rat and Lactobacillus casei (Shirota). J Chromatogr B Analyt Technol Biomed Life Sci. 2006, 844 (2): 240–50. PMID 16876490. doi:10.1016/j.jchromb.2006.07.006. 
  3. ^ Greco, W. R. and Hakala, M. T.,. Evaluation of methods for estimating the dissociation constant of tight binding enzyme inhibitors, (PDF). J. Biol. Chem. 1979, 254 (23): 12104–12109 [2015-11-21]. PMID 500698. (原始内容 (PDF)存档于2009-03-20). 
  4. ^ Dowd, John E.; Riggs, Douglas S. A Comparison of Estimates of Michaelis–Menten Kinetic Constants from Various Linear Transformations. J. Biol. Chem. 1965, 240 (2): 863–869 [2015-11-21]. (原始内容存档于2015-01-22). 

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双倒数图
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