Overexpression, purification and crystallization of an archaeal DNA ligase from Pyrococcus furiosus.
DNA ligases seal single-strand breaks in double-stranded DNA and their function is essential to maintain the integrity of the genome during various aspects of DNA metabolism, such as replication, excision repair and recombination. DNA-strand breaks are frequently generated as reaction intermediates in these events and the sealing of these breaks depends solely on the proper function of DNA ligase. Crystals of the archaeal DNA ligase from Pyrococcus furiosus were obtained using 6.6%(v/v) ethanol as a precipitant and diffracted X-rays to 1.7 A resolution. They belong to the monoclinic space group P2(1), with unit-cell parameters a = 61.1, b = 88.3, c = 63.4 A, beta = 108.9 degrees. The asymmetric unit contains one ligase molecule.
Nishida
Hirokazu
H
Department of Structural Biology, Biomolecular Engineering Research Institute, 6-2-3 Furuedai, Suita, Osaka 565-0874, Japan.
Tsuchiya
Daisuke
D
Ishino
Yoshizumi
Y
Morikawa
Kosuke
K
D020134:Catalytic Domain; D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry; D011088:DNA Ligases / Q000737:chemistry*; D004058:Diffusion; D004926:Escherichia coli / Q000378:metabolism; D000431:Ethanol / Q000494:pharmacology; D020027:Pyrococcus furiosus / Q000201:enzymology*; D013696:Temperature; D014965:X-Rays
D000431:Ethanol; D004247:DNA; D011088:DNA Ligases