Spectral Study on the Photoreduction of Horseradish Peroxidase under Macromolecular Crowding Conditions

doi:10.7517/j.issn.1674-0475.2015.04.321

Abstract

Abstract:

The ferric Horseradish Peroxidase(HRP) can be photoreduced to ferrous state by photoexcitation in dilute solution. However, these researches neglected the high crowding conditions in cell environment. We applied UV-Vis absorption, synchronous fluorescence and CD spectra to study the photoreduction process of HRP and its dependence on the external environment under macromolecular crowding conditions. The UV-Vis absorption spectrum data showed that the degree of HRP photoreduction was higher in Dextran70 solution when 403 nm monochromatic light, but HRP was not photoreduced in Ficoll70 solution. The photoreduction process of HRP was facilitated in Dextran70 or Ficoll70 solution when 280 nm monochromatic light. In addition, the photoreduction extent was much larger in Ficoll70 solution than that in Dextran70. The optimum temperatures of photoreduction process were 4 ℃ and 24 ℃ in the presence of Dextran70 and Ficoll70, respectively. The optimum concentration of Dextran70 for the photoreduction was 100 g/L and photoreduction extent showed an increase with the rise of Ficoll70 concentration. The CD spectra results illustrated that the secondary structure of protein remained the same after irradiation under macromolecular crowding solutions.

Key words: macromolecular crowding environment, horseradish peroxidase, photoreduction, spectroscopy

CAO Hongyu, WANG Zhen, TANG Qian, ZHENG Xuefang. Spectral Study on the Photoreduction of Horseradish Peroxidase under Macromolecular Crowding Conditions[J]. Imaging Science and Photochemistry, 2015, 33(4): 321-329.

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