title: Crystallization and preliminary X-ray analysis of a RecB-family nuclease from the archaeon Pyrococcus abyssi.
abstract: Nucleases are required to process and repair DNA damage in living cells. One of the best studied nucleases is the RecB protein, which functions in Escherichia coli as a component of the RecBCD enzyme complex that amends double-strand breaks in DNA. Although archaea do not contain the RecBCD complex, a RecB-like nuclease from Pyrococcus abyssi has been cloned, expressed and purified. The protein was crystallized by the sitting-drop vapour-diffusion method using polyethylene glycol 8000 as the precipitant. The crystals belong to the orthorhombic space group C222(1), with unit-cell parameters a = 81.5, b = 159.8, c = 100.8 A. Self-rotation function and native Patterson map calculations revealed that there is a dimer in the asymmetric unit with its local twofold axis running parallel to the crystallographic twofold screw axis. The crystals diffracted to about 2 A and a complete native data set was collected to 2.65 A resolution.
authors.affiliation: Center for Structural Biochemistry, Karolinska Institute, NOVUM, S-141 57 Huddinge, Sweden. ren@csb.ki.se
authors.forename: Bin
authors.initials: B
authors.lastname: Ren
authors.forename: Joëlle
authors.initials: J
authors.lastname: Kuhn
authors.forename: Laurence
authors.initials: L
authors.lastname: Meslet-Cladiere
authors.forename: Hannu
authors.initials: H
authors.lastname: Myllykallio
authors.forename: Rudolf
authors.initials: R
authors.lastname: Ladenstein
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D005092:Exonucleases / Q000737:chemistry*; D011487:Protein Conformation; D043903:Pyrococcus abyssi / Q000201:enzymology*
chemical_list: D005092:Exonucleases