title: Expression, purification and crystallization of Helicobacter pylori L-asparaginase.
abstract: The L-asparaginases from Escherichia coli and Erwinia chrysanthemi are effective drugs that have been used in the treatment of acute childhood lymphoblastic leukaemia for over 30 years. However, despite their therapeutic potential, they can cause serious side effects as a consequence of their intrinsic glutaminase activity, which leads to L-glutamine depletion in the blood. Consequently, new asparaginases with low glutaminase activity, fewer side effects and high activity towards L-asparagine are highly desirable as better alternatives in cancer therapy. L-Asparaginase from Helicobacter pylori was overexpressed in E. coli and purified for structural studies. The enzyme was crystallized at pH 7.0 in the presence of 16-19%(w/v) PEG 4000 and 0.1 M magnesium formate. Data were collected to 1.6 A resolution at 100 K from a single crystal at a synchrotron-radiation source. The crystals belong to space group I222, with unit-cell parameters a = 63.6, b = 94.9, c = 100.2 A and one molecule of L-asparaginase in the asymmetric unit. Elucidation of the crystal structure will provide insight into the active site of the enzyme and a better understanding of the structure-activity relationship in L-asparaginases.
authors.affiliation: Turku Centre for Biotechnology, University of Turku and Abo Akademi, Turku 20521, Finland.
authors.forename: Prathusha
authors.initials: P
authors.lastname: Dhavala
authors.forename: Julya
authors.initials: J
authors.lastname: Krasotkina
authors.forename: Christine
authors.initials: C
authors.lastname: Dubreuil
authors.forename: Anastassios C
authors.initials: AC
authors.lastname: Papageorgiou
mesh_terms: D001215:Asparaginase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D016480:Helicobacter pylori / Q000201:enzymology*; D011487:Protein Conformation
chemical_list: D017931:DNA Primers; D001215:Asparaginase