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 Å resolution. All X-ray diffraction data sets are suitable for the determination of the three-dimensional structure of the enzyme.