Crystallization of the 43 kDa ATPase domain of Thermus thermophilus gyrase B in complex with novobiocin.
The 43 kDa ATPase domain of Thermus thermophilus gyrase B was overproduced in Escherichia coli and a three-step purification protocol yielded large quantities of highly purified enzyme which remained stable for weeks. Crystals of the 43 kDa domain in complex with novobiocin, one of the most potent inhibitors of bacterial topoisomerases, were obtained. Crystals obtained in the presence of PEG 8000 do not diffract, but a different crystal form was obtained using sodium formate as a precipitating agent. The plate-shaped crystals, which were less than 10 microm in thickness, could be cryocooled directly from the mother liquor and a full diffraction data set was collected to 2.3 A allowing the determination of the first structure of a gyrase B 43K domain in complex with a coumarin.
Institut de Génétique et de Biologie Moléculaire, CNRS/INSERM/ULP, BP 10142, 1 Rue Laurent Fries, 67404 Illkirch CEDEX, France.
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V
Lamour
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Hoermann
J M
JM
Jeltsch
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Oudet
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D
Moras
D000251:Adenosine Triphosphatases / Q000737:chemistry; D003460:Crystallization; D018360:Crystallography, X-Ray; D027081:DNA Gyrase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D046911:Macromolecular Substances; D008970:Molecular Weight; D009675:Novobiocin / Q000737:chemistry; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics
D046911:Macromolecular Substances; D011994:Recombinant Proteins; D009675:Novobiocin; D000251:Adenosine Triphosphatases; D027081:DNA Gyrase