Cloning, purification and crystallization of a Walker-type Pyrococcus abyssi ATPase family member

Muriel Uhring (1) , Gilbert Bey (1) , Odile Lecompte (1) , Jean Cavarelli (1) , Dino Moras (1) , Olivier Poch (1)

1 IGBMC - Institut de génétique et biologie moléculaire et cellulaire

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.

Domaines: Cristallographie

Muriel Uhring, Gilbert Bey, Odile Lecompte, Jean Cavarelli, Dino Moras, et al.. Cloning, purification and crystallization of a Walker-type Pyrococcus abyssi ATPase family member. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 2005, 61 (Pt 10), pp.925-927. ⟨10.1107/S174430910502868X⟩. ⟨hal-04122991⟩