期刊
IUCRJ
卷 7, 期 -, 页码 1084-1091出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S2052252520012142
关键词
mesophase crystallization; lipid sponge phase; monoolein; photosynthetic reaction centers; Rhodobacter sphaeroides; serial crystallography
资金
- program of the Presidium of the Russian Academy of Sciences 'Molecular and Cell Biology and Postgenomic Technologies'
- Russian Foundation for Basic Research [18-02-40008, 17-00-00207]
- Cluster of Excellence 'Advanced Imaging of Matter' of the Deutsche Forschungsgemeinschaft (DFG) [EXC 2056, 390715994]
- BMBF [50WB1422, 05K16GUA]
- Deutsche Forschungsgemeinschaft [390715994]
- Bundesministerium fur Bildung und Forschung [50WB1422, 05K16GUA]
With the recent developments in the field of free-electron-laser-based serial femtosecond crystallography, the necessity to obtain a large number of highquality crystals has emerged. In this work crystallization techniques were selected, tested and optimized for the lipid mesophase crystallization of the Rhodobacter sphaeroides membrane pigment-protein complex, known as the photosynthetic reaction center (RC). Novel approaches for lipid sponge phase crystallization in comparatively large volumes using Hamilton gas-tight glass syringes and plastic pipetting tips are described. An analysis of RC crystal structures obtained by lipid mesophase crystallization revealed non-native ligands that displaced the native electron-transfer cofactors (carotenoid spheroidene and a ubiquinone molecule) from their binding pockets. These ligands were identified and were found to be lipids that are major mesophase components. The selection of distinct co-crystallization conditions with the missing cofactors facilitated the restoration of spheroidene in its binding site.
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