Fluorescence photobleaching recovery measurement of protein absolute diffusion constants

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...

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Bibliographic Details
Published in:Biophysical chemistry 1979-09, Vol.10 (2), p.221-229
Main Authors: Barisas, B.George, Leuther, Michael D.
Format: Article
Language:English
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Summary: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.
ISSN:0301-4622
1873-4200
DOI:10.1016/0301-4622(79)85044-9