title: Crystallization and preliminary crystallographic analysis of L-asparaginase from Erwinia carotovora.
abstract: Bacterial L-asparaginases have been used as therapeutic agents in the treatment of acute childhood lymphoblastic leukaemia for over 30 y. However, their use is limited owing to the glutaminase activity of the administered enzymes, which results in serious side effects. In contrast, L-asparaginase from Erwinia carotovora exhibits low glutaminase activity at physiological concentrations of L-asparagine and L-glutamine in the blood. Recombinant Er. carotovora L-asparaginase was crystallized in the presence of L-glutamate by the hanging-drop vapour-diffusion method using 10 mg ml(-1) purified enzyme, 16-18%(w/v) PEG 3350 and 0.2 M NaF. X-ray diffraction data were collected to 2.6 A at 293 K using an in-house rotating-anode generator. The crystals belong to the monoclinic P2(1) space group, with unit-cell parameters a = 78.0, b = 112.3, c = 78.7 A, beta = 101.9 degrees and a homotetramer in the crystallographic asymmetric unit. A molecular-replacement solution has been found and refinement is currently in progress. The crystal structure may provide leads towards protein-engineering efforts aimed at safer asparaginase administration in leukaemia treatment.
authors.affiliation: Turku Centre for Biotechnology, University of Turku, Turku 20521, Finland.
authors.forename: Linnea E K
authors.initials: LE
authors.lastname: Wikman
authors.forename: Julya
authors.initials: J
authors.lastname: Krasotkina
authors.forename: Anastasia
authors.initials: A
authors.lastname: Kuchumova
authors.forename: Nikolay N
authors.initials: NN
authors.lastname: Sokolov
authors.forename: Anastassios C
authors.initials: AC
authors.lastname: Papageorgiou
mesh_terms: D001215:Asparaginase / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D003460:Crystallization / Q000379:methods; D018698:Glutamic Acid / Q000737:chemistry; D016973:Pectobacterium carotovorum / Q000737:chemistry*; D011092:Polyethylene Glycols; D011994:Recombinant Proteins; D014835:Volatilization; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D018698:Glutamic Acid; D011092:Polyethylene Glycols; D001215:Asparaginase