Kinetics of denaturation of the yellow fluorescent protein from coral zFP538

Nadya N. Zubova; Vadim A. Korolenko; Alexander P. Savitsky

doi:10.1117/12.590272

4 April 2005 Kinetics of denaturation of the yellow fluorescent protein from coral zFP538

Nadya N. Zubova, Vadim A. Korolenko, Alexander P. Savitsky

Proceedings Volume 5704, Genetically Engineered and Optical Probes for Biomedical Applications III; (2005) https://doi.org/10.1117/12.590272
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

Our experiments show that fluorescence dependence of zFP538 on protein concentration is nonlinear over the range from 1.2•10^-9 M to 5.5•10^-7 M. Under pH from 5 to 9 it can be divided into two linear regions with different slope at concentration more and less than 1•10^-7 M. One may conclude, there are at least two different forms of the yellow protein observed in the high (>1•10^-7 M) and low (<1•10^-7 M) concentration regions. These forms may be different aggregation states of the protein. We have obtained acid denaturation kinetics curves for zFP538 over pH range from 3.5 to 5.5 at different protein concentrations. Under all pH values relative residual fluorescence of the most concentrated solution (1.33•10^-6 M) is higher than that for the solutions of lower protein concentration (1.36•10^-7 M and 1.36•10^-8 M). Fluorescence decrease under pH from 3.5 to 4.24 is three or two exponential. Under higher pH values kinetic curves have a plateau at first and then fluorescence drops in exponential way. The results obtained support the hypothesis that at higher protein concentrations zFP538 is more stable against acid denaturation.

Citation Download Citation

Nadya N. Zubova, Vadim A. Korolenko, and Alexander P. Savitsky "Kinetics of denaturation of the yellow fluorescent protein from coral zFP538", Proc. SPIE 5704, Genetically Engineered and Optical Probes for Biomedical Applications III, (4 April 2005); https://doi.org/10.1117/12.590272

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