Protein crystallization studies – complex overview

Ivana Kuta Smatanova^{1, 2}

¹ University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses and School of complex systems, Zamek 136, 373 33 Nove Hrady

² Academy of Sciences of the Czech Republic, Inst. of Nanobiology and Structural Biology GCRC, Zamek 136, 373 33 Nove Hrady

The research at the Laboratory of crystallogenesis and biomolecular crystallography of Institute of Nanobiology and Structural Biology GCRC AS CR joined with Laboratory of molecular structure and dynamics of School of complex systems of FFPW University of South Bohemia in Ceske Budejovice is aimed at crystallization and crystallographic studies of soluble and membrane proteins and protein complexes. Since y. 2000 when the research in this field has begun, more than 20 proteins, protein complexes and mutant variants such as HsdR subunit of restriction enzyme of EcoR124I from E.coli plus its 3 mutant forms, PsbP protein from Spinacia oleracea, WrbA apo and holo forms of E.coli, Fe-regulated protein D (FrpD) of Neisseria meningitides, haloalkan dehalogenases DhaAwt, DhaA04, DhaA12, DhaA13, DhaA14, DhaA15, DhaA31 from Rhodococcus rhodochrous NCIMB 13064 and their complexes with substrates isopropanol (IPA) and 1, 2, 3 – trichloropropane (TCP), DbeA and DbeA1 of Bradyrhizobium elkani USDA94 as well as two new haloalkan dehalogenases DpcA from Psychrobacter cryohalolentis K5 and DmxA from Marynobacter sp ELB 17 were crystallized, tested and crystals were measured at the synchrotron radiation sources. Diffraction data were used to solve and refine protein structures that have been deposited in PDB under specific codes. Almost all enzymes were crystallized using standard, advanced and alternative crystallization techniques e.g. cross influence procedure that was directly developed and tested in our laboratory.

This research was supported by the ME CR (COST LD11011, ME09016, CZ.1.05/2.1.00/01.0024), GACR (P207/12/0775, P207/11/0717), by the AS CR (AV0Z60870520).