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.
Institute of Agricultural Chemistry, University of Bologna, Viale Berti Pichat 10 I-40127 Bologna, Italy.
S
S
Benini
S
S
Ciurli
W R
WR
Rypniewski
K S
KS
Wilson
S
S
Mangani
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