Crystallization and preliminary high-resolution X-ray diffraction analysis of native and beta-mercaptoethanol-inhibited urease from Bacillus pasteurii.
Hexagonal crystals of urease from Bacillus pasteurii have been obtained by vapour diffusion at 293 K in 20 mM Tris-HCl, neutral pH, containing 50 mM Na2SO3. Isomorphous crystals of urease inhibited with beta-mercaptoethanol were also obtained by including 4 mM of the inhibitor in the enzyme solution. Crystals of the native and inhibited enzyme diffract respectively to 2.00 A (96.7% completeness) and to 1.65 A (98.7% completeness) using synchrotron X-ray cryogenic (100 K) conditions. The space group is P6322 for both forms, and the unit-cell parameters are a = b = 131.36, c = 189. 76 A for native urease and a = b = 131.34, c = 190.01 A for inhibited urease. Under the same conditions, single crystals of B. pasteurii urease inhibited with acetohydroxamic acid, cisteamine, and phenylphosphorodiamidate were also obtained.
Benini
S
S
Institute of Agricultural Chemistry, University of Bologna, Viale Berti Pichat 10 I-40127 Bologna, Italy.
Ciurli
S
S
Rypniewski
W R
WR
Wilson
K S
KS
Mangani
S
S
D001407:Bacillus / Q000201:enzymology*; D003460:Crystallization; D003543:Cysteamine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006877:Hydroxamic Acids / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology*; D017356:Synchrotrons; D014510:Urease / Q000037:antagonists & inhibitors*; D014961:X-Ray Diffraction
D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; C006358:acetohydroxamic acid; D003543:Cysteamine; D008623:Mercaptoethanol; D014510:Urease