Open AccessCrystallization and preliminary crystallographic analysis of the tetrameric form of phosphofructokinase‐2 from Escherichia coli , a member of the ribokinase family
Author(s) -
Cabrera Ricardo,
Caniuguir Andrés,
Ambrosio Andre L. B.,
Guixé Victoria,
Garratt Richard C.,
Babul Jorge
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106032246
Subject(s) - escherichia coli , crystallization , crystallography , phosphofructokinase , chemistry , genetics , biology , biochemistry , enzyme , gene , glycolysis , organic chemistry
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 P 222 1 , with unit‐cell parameters a = 42.8, b = 86.8, c = 171.3 Å. The calculated Matthews coefficient of 2.45 Å 3 Da −1 indicates the presence of two monomers in the asymmetric unit, corresponding to a solvent content of 49%. Structure determination is ongoing.