Download MendeleyExpression, purification, crystallization and preliminary crystallographic analysis of a thermostable DNA ligase from the archaeon Thermococcus sibiricus.
Petrova, T.E., Bezsudnova, E.Y., Dorokhov, B.D., Slutskaya, E.S., Polyakov, K.M., Dorovatovskiy, P.V., Ravin, N.V., Skryabin, K.G., Kovalchuk, M.V., Popov, V.O.(2012) Acta Crystallogr Sect F Struct Biol Cryst Commun 68: 163-165
- PubMed: 22297989
- DOI: https://doi.org/10.1107/S1744309111050913
- Primary Citation of Related Structures:
4EQ5 - PubMed Abstract:
DNA ligases join single-strand breaks in double-stranded DNA by catalyzing the formation of a phosphodiester bond between adjacent 5'-phosphate and 3'-hydroxyl termini. Their function is essential to maintain the integrity of the genome in DNA replication, recombination and repair. A recombinant ATP-dependent DNA ligase from the hyperthermophilic anaerobic archaeon Thermococcus sibiricus was expressed in Escherichia coli and purified. Crystals were grown by vapour diffusion using the hanging-drop method with 17%(w/v) PEG 4000 and 8.5%(v/v) 2-propanol as precipitants. A diffraction experiment was performed with a single crystal, which diffracted X-rays to 3.0 Å resolution. The crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 58.590, b = 87.540, c = 126.300 Å.
Organizational Affiliation:
Bach Institute of Biochemistry, RAS, Moscow, Russian Federation. petrova@impb.psn.ru
