Crystal engineering: a case study using the 24 kDa fragment of the DNA gyrase B subunit from Escherichia coli.
Site-directed mutagenesis was used to determine the efficacy of changing surface residues to improve crystal quality. Nine mutants of the 24 kDa fragment of the Escherichia coli DNA gyrase B subunit were produced, changing residues on the protein's surface. The mutations changed either the charge or the polarity of the wild-type amino acid. It was found that single amino-acid changes on the surface could have a dramatic effect on the crystallization properties of the protein and generally resulted in an improvement in the number of crystal-screen hits as well as an improvement in crystal quality. It is concluded that crystal engineering is a valuable tool for protein crystallography.
D'Arcy
A
A
F. Hoffmann-La Roche Ltd Pharmaceutical Research, Chemical Technologies, CH-4070, Basel, Switzerland.
Stihle
M
M
Kostrewa
D
D
Dale
G
G
D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology*; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry*; D015203:Reproducibility of Results
D001426:Bacterial Proteins; D010446:Peptide Fragments; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II