Preliminary crystallographic studies of Y25F mutant of periplasmic Escherichia coli L-asparaginase.

• Authors: Maciej Kozak , Mariusz Jaskólski , Klaus H Röhm
• Languages of publication: EN

Abstracts

EN

Periplasmic Escherichia coli L-asparaginase II with Y25F mutation in the active-site cavity has been obtained by recombinant techniques. The protein was crystallized in a new hexagonal form (P6522). Single crystals of this polymorph, suitable for X-ray diffraction, were obtained by vapor diffusion using 2-methyl-2,4-pentanediol as precipitant (pH 4.8). The crystals are characterized by a = 81.0, c = 341.1 Å and diffract to 2.45 Å resolution. The asymmetric unit contains two protein molecules arranged into an AB dimer. The physiologically relevant ABA'B' homotetramer is generated by the action of the crystallographic 2-fold axis along [1, -1, 0]. Kinetic studies show that the loss of the phenolic hydroxyl group at position 25 brought about by the replacement of Y with F strongly impairs kcat without significantly affecting Km.

Keywords

EN

protein crystallography; mutagenesis; leukemia; amidohydrolase

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2000
• Volume: 47
• Issue: 3
• Pages: 807-814

Dates

published

2000

received

2000-05-12

accepted

2000-06-19

Contributors

author

Maciej Kozak

• Department of Macromolecular Physics, Faculty of Physics, A. Mickiewicz University, Poznań, Poland

author

Mariusz Jaskólski

• Department of Crystallography, Faculty of Chemistry, A. Mickiewicz University, Poznań, Poland

author

Klaus H Röhm

• Institut für Physiologische Chemie, Philipps Universität, Marburg, Germany

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