| Measurement of mouse liver glutathione S-transferase activity by the integrated method |
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| 引用本文： | 廖飞,李甲初,康格非,曾昭淳,左渝萍. Measurement of mouse liver glutathione S-transferase activity by the integrated method[J]. 中国人民解放军军医大学学报, 2003, 18(5): 295-300 |
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| 作者姓名： | 廖飞 李甲初 康格非 曾昭淳 左渝萍 |
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| 作者单位： | Department of Biochemistry，Chongqing University of Medical Sciences，Chongqing 400016，China，Department of Biochemistry，Chongqing University of Medical Sciences，Chongqing 400016，China，Faculty of Laboratory Medicine，Chongqing University of Medical Sciences，Chongqing 400016，China，Department of Biochemistry Chongqing University of Medical Sciences，Chongqing 400016，China，Department of Biochemistry Chongqing University of Medical Sciences，Chongqing 400016，China |
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| 基金项目： | National Natural Science Foundation of China (No.30200266) |
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| 摘 要： | Objective: The integrated method was investigated to measure Vm/Km of mouse liver glutathione S-transfer-ase (GST) activity on GSH and 7-Cl-4-nitrobenzofurazozan. Methods: Presetting concentration of one substrate twenty-fold above the other's and taking maximum product absorbance Am as parameter while Km as constant, Vm/Km was obtained by nonlinear fitting of GST reaction curve to the integrated Michaelis-Menten equation In [Am/(Am -Ai)] + Ai/ ( ξ× Km ) = ( Vm/Km )×ti (1). Results: Vm/Km for GST showed slight dependence on initial substrate concentration and data range, but it was resistant to background absorbance, error in reaction origin and small deviation in presetting Km. Vm/Km was proportional to the amount of GST with upper limit higher than that by initial rate. There was close correlation between Vm/Km and initial rate of the same GST. Consistent results were obtained by this integrated method and classical initial rate method for the measurement of mouse liver GST. Conclusion: With the co
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| 关 键 词： | 肝脏 谷胱甘肽S-转移酶 Vm Km |
Measurement of mouse liver glutathione S-transferase activity by the integrated method |
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| LIAO Fei LI Jia-chu,KANG Ge-fei ZENG Zhao-chun ZUO Yu-ping. Measurement of mouse liver glutathione S-transferase activity by the integrated method[J]. Journal of Medical Colleges of PLA(China), 2003, 18(5): 295-300 |
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| Authors: | LIAO Fei LI Jia-chu KANG Ge-fei ZENG Zhao-chun ZUO Yu-ping |
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| Affiliation: | LIAO Fei LI Jia-chu,KANG Ge-fei ZENG Zhao-chun ZUO Yu-ping Department of Biochemistry,Faculty of Laboratory Medicine,Chongqing University of Medical Sciences,Chongqing400016,China |
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| Abstract: | Objective: The integrated method was investigated to measure Vm/Km of mouse liver glutathione S-transferase (GST) activity on GSH and 7-Cl-4-nitrobenzofurazozan. Methods: Presetting concentration of one substrate twenty-fold above the others and taking maximum product absorbance Am as parameter while Km as constant, Vm/Km was obtained by nonlinear fitting of GST reaction curve to the integrated Michaelis-Menten equation ln [Am/(Am-Ai)]+Ai/(ε×Km)=(Vm/Km)×ti (1). Results: Vm/Km for GST showed slight dependence on initial substrate concentration and data range, but it was resistant to background absorbance, error in reaction origin and small deviation in presetting Km. Vm/Km was proportional to the amount of GST with upper limit higher than that by initial rate. There was close correlation between Vm/Km and initial rate of the same GST. Consistent results were obtained by this integrated method and classical initial rate method for the measurement of mouse liver GST. Conclusion: With the concentration of one substrate twenty-fold above the others, this integrated method was reliable to measure the activity of enzyme on two substrates, and substrate concentration of the lower one close to its apparent Km was able to be used. |
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| Keywords: | integrated method glutathione S-transferase nonlinear fitting reaction curve predictor variable enzyme activity measurement |
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