ИССЛЕДОВАНИЕ ФОТОХИМИЧЕСКИХ СВОЙСТВ И ФИЛОГЕНИИ РАСТИТЕЛЬНЫХ БЕЛКОВ СЕМЕЙСТВА WSCP
Here we show the ability of the chlorophyll associated with proteins of the WSCP family (Water-Soluble Chlorophyll-binding Proteins) to photosensitize oxidative redox reactions. Irradiation with red light of the recombinant chlorophyll-protein complexes WSCP subclasses IIa and IIb in the presence of an electron donor (NADH) led to oxidation of the donor, i.e., these pigment-protein complexes showed photochemical activity. Meanwhile there was no photodestruction of chlorophyll associated with WSCP, which indicates the photocatalytic nature of the detected redox reaction. The kinetic constants of NADH photooxidation were higher for WSCP subclass IIa (BoWSCP) than for subclass IIb (LvWSCP). To explain the difference in the photosensitizing activity of representatives of different WSCP subclasses, bioinformatic analysis of class II proteins of this family was carried out. For this purpose, we searched for members of the WSCP family in the UniProt protein sequence database using the BLAST search algorithm, followed by their multiple alignment and construction of a phylogenetic tree using the EMBL-EBI Clustal Omega web service and the MEGA7 program. Bioinformatic analysis has confirmed the phylogenetic division of the WSCP class II protein family into two subclasses, previously established on the basis of the difference in their physicochemical properties. It was suggested that phylogeny is related to the photochemical activity of representatives of different subclasses in the WSCP family.
2. Hirabayashi H, Amakawa M, Kamimura Y, et al. Analysis of photooxidized pigments in watersoluble chlorophyll protein complex isolated from Chenopodium album. J. Photochem. Photobiol. A.
3.Satoh H, Uchida A, Nakayama K, et al. Watersoluble chlorophyll protein in Brassicaceae plants is a stress-induced chlorophyll-binding protein. Plant Cell
Physiol. 2001;42(9):906-911. https://doi.org/10.1093/pcp/pce117
4.Takahashi S, Yanai H, Oka‑Takayama Y, et al. Molecular cloning, characterization and analysis of the intracellular localization of a water-soluble chlorophyll-binding protein (WSCP) from Virginia pepperweed (Lepidium virginicum), a unique WSCP that preferentially binds chlorophyll b in vitro. Planta. 2013;238(6):1065-1080. https://doi.org/10.1007/s00425-013-1952-7
5. Rustgi S, Boex-Fontvieille E, Reinbothe C, et al. Serpin1 and WSCP differentially regulate the activity of the cysteine protease RD21 during plant development in Arabidopsis thaliana. Proc. Natl. Acad. Sci. U. S. A. 2017;114(9):2212-2217. https://doi.org/10.1073/pnas.1621496114
6.Horigome D, Satoh H, Itoh N, et al. Structural mechanism and photoprotective function of watersoluble chlorophyll-binding protein. J. Biol. Chem.
7.Bednarczyk D, Dym O, Prabahar V et al. Fine tuning of chlorophyll spectra by protein‐induced ring deformation. Angew. Chem., Int. Ed. Engl. 2016;55(24):6901-6905. https://doi.org/10.1002/anie.201512001
8.Palm D M, Agostini A, Tenzer S, et al. Watersoluble chlorophyll protein (WSCP) stably binds two or four chlorophylls. Biochemistry. 2017;56(12):1726-
1736. https://doi.org/10.1021/acs.biochem.7b00075 9.Takahashi S, Yanai H, Nakamaru Y, et al. Molecular cloning, characterization and analysis of the intracellular localization of a water-soluble Chlbinding protein from Brussels sprouts (Brassica oleracea var. gemmifera). Plant Cell Physiol. 2012;53(5):879-891. https://doi.org/10.1093/pcp/pcs031
10.Schmidt K, Fufezan C, Krieger-Liszkay A, et al. Recombinant water-soluble chlorophyll protein from Brassica oleracea var. Botrys binds various chlorophyll derivatives. Biochemistry. 2003;42(24):7427-7433. https://doi.org/10.1021/bi034207r
11. Charlton A, Zachariou M. Immobilized metal ion affinity chromatography of histidine-tagged fusion proteins. In Affinity Chromatography. Humana Press.
12. Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal.
Biochem. 1976;72(1-2):248-254. https://doi.org/10.1016/0003-2697(76)90527-3