Escherichia coli B γ-glutamylcysteine synthetase:: modification, purification, crystallization and preliminary crystallographic analysis

Escherichia coli B gamma-glutamylcysteine synthetase (gammaGCS) catalyzes the ATP-dependent coupling of L-Glu and L-Cys to form the glutathione precursor gamma-L-Glu-Cys and is a target for development of potential therapeutic agents. By introducing four point mutations of surface-exposed cysteine residues to serine, the gammaGCS was purified to homogeneity; single crystals have been obtained using the hanging-drop vapour-diffusion method with sodium formate. The GCS crystal diffracted to 2.8 Angstrom and belongs to space group R3, with unit-cell parameters a = b = 326.7, c = 103.9 Angstrom.