Crystallization and preliminary crystallographic analysis of the tetrameric form of phosphofructokinase-2 from Escherichia coli, a member of the ribokinase family
- Laboratorio de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Santiago (Chile)
- Centro de Biotecnologia Molecular Estrutural, Instituto de Física de São Carlos, Universidade de São Paulo, Avenida Trabalhador Sãocarlense 400, CEP 13560-970, São Carlos, SP (Brazil)
The phosphofructokinase-2 enzyme from E. coli was crystallized in its tetrameric inhibited form. This is the only member of the ribokinase family known to suffer a transition from dimer to tetramer in response to the allosteric binding of MgATP. Escherichia coli contains two phosphofructokinases, Pfk-1 and Pfk-2, which belong to unrelated protein families. In addition to catalytic function, the enzymes have converged in showing substrate inhibition by the nucleotide MgATP. However, although both Pfk-1 and Pfk-2 have been extensively characterized biochemically, only the structure of the former has been solved by X-ray diffraction. In order to fully understand how the same function has evolved on different structural folds, Pfk-2 has been crystallized by the hanging-drop vapour-diffusion method using PEG 6000 as precipitant. Single crystals were grown in the presence of MgATP and diffracted to 1.98 Å. The crystals belong to the orthorhombic system, space group P222{sub 1}, with unit-cell parameters a = 42.8, b = 86.8, c = 171.3 Å. The calculated Matthews coefficient of 2.45 Å{sup 3} Da{sup −1} indicates the presence of two monomers in the asymmetric unit, corresponding to a solvent content of 49%. Structure determination is ongoing.
- OSTI ID:
- 22360154
- Journal Information:
- Acta Crystallographica. Section F, Journal Name: Acta Crystallographica. Section F Journal Issue: Pt 9 Vol. 62; ISSN ACSFCL; ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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