Crystallization, structural and modeling studies of photosynthetic membrane protein and water soluble haloalkane dehalogenase

PRUDNIKOVA, Tatyana

This PhD thesis is devoted to study of a) water soluble protein haloalkane dehalogenase DbeA (wild type enzyme) and its mutant variant DbeA1 from bacteria Bradyrhizobium elkani USDA94 and also b) the inner membrane protein complex of Photosystem II (PSII) from Pisum sativum L. An active site architecture of the haloalkane dehalogenase enzymes DbeA, DbeA1, stabilization by loop insertion in a mutant form, and structure-function relationships of haloalkane dehalogenases have been studied by using experimental approaches protein crystallization and crystallography. The protein crystals have been grown and X-ray diffraction data on these crystals have been collected and 3D molecular structure has been solved for both dehalogenases DbeA and DbeA1 at 2.2 A? resolution, for DbeA crystals soaked with 1-fluoropentane as a substrate at 1.65 A? resolution, and with 1,3-dichloropropane substrate at 2.15 A? resolution. Crystallization and molecular modelling methods were applied on membrane protein PSII core complex from higher plants in order to find optimal conditions for the 3D crystals formation, to establish the crystallization protocol and to study light-induced processes in PSII reaction center.

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