Crystallization and Preliminary X-ray Diffraction Study of a Mutant of L-Asparaginase from Wolinella succinogenes

Abstract

The double mutant of Wolinella succinogenes L-asparaginase (Was72) with the V23Q and K24T substitutions in the C-terminal region of the N-terminal loop enclosing the active site was crystallized in the apo form and in complexes with L-aspartic and L-glutamic acids. This mutant exhibits glutaminase activity eight times lower compared to the wild-type enzyme. Crystals of the apo enzyme were grown in two modifications (sp. gr. P21 and sp. gr. P22121). Crystals grown in the presence of both aspartic and glutamic acids belong to the same space group (P21) but have different unit cell parameters. The X-ray diffraction data sets were collected from all types of crystals at 1.65–2.00 A resolution. All X-ray diffraction data sets are suitable for the determination of the three-dimensional structure of the enzyme.