A Convenient Method for Determination of Dissociation Constants of Enzyme-Ligand Complexes Based on Difference Absorption Spectra. : A Model Case of Ribonuclease F1-Guanosine 2'-Monophosphate
書誌事項
- タイトル別名
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- Convenient Method for Determination of
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A convenient method has been developed for investigating interaction of an enzyme with a specific ligand on the basis of the difference absorption spectrum. The experimental procedure has been described in detail. Briefly, to a solution of the enzyme, a concentrated solution of the ligand was added successively in small volume aliquots and the increase in absorbance after each addition was measured at two wavelengths which corresponded to a maximum and a crossover point of the difference absorption spectrum. Processing the obtained data gave the difference at the maximum wavelength as a function of the total ligand concentration. The data set thus generated was fitted to the equilibrium equation and the best fit values for the dissociation constant, K_d and the molar difference absorption coefficient at the maximum wavelength, Aemax, were determined. The method was successfully applied to ribonuclease F1-guanosine 2'-phosphate system and gave the following parameters: K_d=2.5 μM and Δε_<290>=3890 M^<-1>cm^<-1>.
収録刊行物
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- Bulletin of Shimane Medical University
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Bulletin of Shimane Medical University 18 93-96, 1995-12-01
島根医科大学
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詳細情報 詳細情報について
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- CRID
- 1050845763416196224
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- NII論文ID
- 110000979051
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- NII書誌ID
- AN00107602
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- ISSN
- 03879097
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- NDL書誌ID
- 3956261
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- 本文言語コード
- en
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- 資料種別
- departmental bulletin paper
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- データソース種別
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- IRDB
- NDL
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