title: Cloning, purification and crystallization of a Walker-type Pyrococcus abyssi ATPase family member.
abstract: Several ATPase proteins play essential roles in the initiation of chromosomal DNA replication in archaea. Walker-type ATPases are defined by their conserved Walker A and B motifs, which are associated with nucleotide binding and ATP hydrolysis. A family of 28 ATPase proteins with non-canonical Walker A sequences has been identified by a bioinformatics study of comparative genomics in Pyrococcus genomes. A high-throughput structural study on P. abyssi has been started in order to establish the structure of these proteins. 16 genes have been cloned and characterized. Six out of the seven soluble constructs were purified in Escherichia coli and one of them, PABY2304, has been crystallized. X-ray diffraction data were collected from selenomethionine-derivative crystals using synchrotron radiation. The crystals belong to the orthorhombic space group C2, with unit-cell parameters a = 79.41, b = 48.63, c = 108.77 A, and diffract to beyond 2.6 A resolution.
authors.affiliation: Département de Biologie et Génomiques Structurales, UMR 7104, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP Strasbourg, 1 Rue Laurent Fries, 64404 Illkirch, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Uhring
authors.forename: Gilbert
authors.initials: G
authors.lastname: Bey
authors.forename: Odile
authors.initials: O
authors.lastname: Lecompte
authors.forename: Jean
authors.initials: J
authors.lastname: Cavarelli
authors.forename: Dino
authors.initials: D
authors.lastname: Moras
authors.forename: Olivier
authors.initials: O
authors.lastname: Poch
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D020816:Amino Acid Motifs; D019843:Archaeal Proteins / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology; D004247:DNA / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D005822:Genetic Vectors; D023281:Genomics; D015233:Models, Statistical; D011487:Protein Conformation; D043903:Pyrococcus abyssi / Q000201:enzymology*; D012645:Selenomethionine / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D019843:Archaeal Proteins; D004247:DNA; D012645:Selenomethionine; D000251:Adenosine Triphosphatases