FLUORESCENCE PHOTOBLEACHING RECOVERY MEASUREMENT OF PROTEIN ABSOLUTE DIFFUSION CONSTANTS

被引:60
作者
BARISAS, BG
LEUTHER, MD
机构
[1] Edward A. Doisy Department of Biochemistry, St. Louis University School of Medicine, St. Louis
基金
美国国家科学基金会;
关键词
D O I
10.1016/0301-4622(79)85044-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The technique of fluorescence photobleaching recovery [Axelrod et al., Biophys. J. 16 (1976) 1055] has been applied to the measurement of absolute diffusion constants of a number of fluoiescein isothiocyanate-labeled proteins. Measured diffusion constants agree to within ± 7% of published values for the underivatized proteins. The method has sufficient sensitivity to reveal the concentration dependence at neutral pH of the diffusion constant of α-chymotrypsin. The rapidity with which the labelling and measurements can be performed and the small amount of material required suggest the technique may be useful in rapid characterization of small protein samples. Some developments in optical and electronic systems and in data processing for this technique are discussed. © 1979.
引用
收藏
页码:221 / 229
页数:9
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