EC Number   |
Reference |
|---|
    1.3.1.98 | - |
286070 |
| 1.3.1.98 | by sitting-drop vapour diffusion |
286071 |
| 1.3.1.98 | enzyme bound to FAD and K+, hanging drop vapor diffusion method |
765472 |
| 1.3.1.98 | purified recombinant enzyme MurB complexed with FAD and NADP+ in two crystal forms, hanging drop vapour diffusion method, for crystal type A: mixing of 0.001 ml of protein solution containing 25 mg/ml PaMurB, and 2 mM unbuffered NADPH, with 0.001 ml of reservoir solution containing 0.1 M Bis-Tris propane, pH 7.0, 0.2 M sodium potassium tartrate, and 15% w/v PEG 3350, and equilibration against 0.5 ml reservoir solution, crystals from these drops are cryoprotected with a solution containing additional 15% w/v PEG3350 and 2 mM NADPH before flash freezing, for crystal type B: the protein is mixed with a reservoir solution containing 40 mM potassium phosphate, 20% v/v glycerol, 16% w/v PEG 8000 and 2 mM Tris-buffered NADP+ sodium salt, the crystals are frozen directly without addition of a cryoprotectant, X-ray diffraction structure determination and analysis at 2.2 A and 2.1 A resolution, respectively, molecular replacement using the atomic coordinates of the complex of Escherichia coli MurB with UDP-N-acetylglucosamine enolpyruvate, PDB code 2MBR, stripped of all ligands and water molecules as search model, modeling |
746242 |
| 1.3.1.98 | purified recombinant His-tagged enzyme, 20 mg/ml protein in 20 mM HEPES, pH 7.5 5 mM 2-mercaptoethanol or DTT, 20°C, hanging drop vapour diffusion method, mixing with reservoir solution containing 0.1 M sodium cacodylate, pH 6.5, 15% PEG 8000, and 0.5 M (NH4)2SO4, 4 days, yellow plate-shaped crystals cryoprotected in 10% PEG 8000, 0.55 M (NH4)2SO4, 0.1 M sodium cacodylate, pH 6.5, 5 mM BME or DTT with 15% 2-methyl 2,4-pentanediol, X-ray diffraction structure determination and analysis at 2.3 A resolution, altered crystallization conditions were optimized using the purified selenomethionine-labeled MurB at 20 mg/ml, in complex with 10 mM enolpyruvyl-UDP-N-acetylglucosamine, in 1-2 weeks on grease-coated microbridges by mixing 5 ml Se-Met MurB and 5 ml reservoir solution containing 0.1 M sodium cacodylate, 9.75% PEG 8000, 0.55 M (NH4)2SO4, 20% DMSO, and 5 mM 2-mercaptoethanol and producing larger crystals, overview |
286071 |
| 1.3.1.98 | purified recombinant MurB, hanging drop-vapor diffusion method, 0.001 ml of protein solution containing 20 mg/ml protein in 20 mM HEPES-NaOH, pH 7.5, is mixed with 0.001 ml of reservoir solution containing 100 mM MES-NaOH, pH 6.5, 18% w/v PEG 8000 and 200 mM CaCl2, equilibration against 1 ml of mother liquor, Se-Met substituted crystals are grown under the same conditions, while the substrate-complex crystals are grown in the presence of 25 mM substrate UDPGlcNAc, crystals appear within 8 h, X-ray difraction structure determination and analysis at 1.15-1.7 A resolution |
689925 |
| 1.3.1.98 | S229A mutant and wild-type MurB |
286069 |
| 1.3.1.98 | sequence alignment with Escherichia coli structures, PDB entries 2Q85 and 2MBR, and modeling of structure as well as docking with N-acetylpyruvyl glucosamine |
698345 |
