A comparative study of three signaling forms of the orange carotenoid protein
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- 作者：E. G. Maksimov ; M. Moldenhauer ; E. A. Shirshin ; E. A. Parshina…
- 关键词：Orange carotenoid protein ; Synechocystis sp. PCC 6803 ; Protein expression ; Echinenone ; Fluorescence lifetime ; Fluorescence anisotropy
- 刊名：Photosynthesis Research
- 出版年：2016
- 出版时间：December 2016
- 年：2016
- 卷：130
- 期：1-3
- 页码：389-401
- 全文大小：1,520 KB
- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Life Sciences
Plant Physiology - 出版者：Springer Netherlands
- ISSN：1573-5079
- 卷排序：130
文摘
Orange carotenoid protein (OCP) is a water-soluble photoactive protein responsible for a photoprotective mechanism of nonphotochemical quenching in cyanobacteria. Under blue–green illumination, OCP converts from the stable orange into the signaling red quenching form; however, the latter form could also be obtained by chemical activation with high concentrations of sodium thiocyanate (NaSCN) or point mutations. In this work, we show that a single replacement of tryptophan-288, normally involved in protein-chromophore interactions, by alanine, results in formation of a new protein form, hereinafter referred to as purple carotenoid protein (PCP). Comparison of resonance Raman spectra of the native photoactivated red form, chemically activated OCP, and PCP reveals that carotenoid conformation is sensitive to the structure of the C-domain, implicating that the chromophore retains some interactions with this part of the protein in the active red form. Combination of differential scanning fluorimetry and picosecond time-resolved fluorescence anisotropy measurements allowed us to compare the stability of different OCP forms and to estimate relative differences in protein rotation rates. These results were corroborated by hydrodynamic analysis of proteins by dynamic light scattering and analytical size-exclusion chromatography, indicating that the light-induced conversion of the protein is accompanied by a significant increase in its size. On the whole, our data support the idea that the red form of OCP is a molten globule-like protein in which, however, interactions between the carotenoid and the C-terminal domain are preserved.