    2.7.1.130 | purified LpxK in complex with the ATP analogue AMPPCP in the closed catalytically competent conformation, sitting drop vapor diffusion method, mixing of 0.70 ml well solution with 0.01 ml sample solution, containing four parts of a reservoir solution consisting of 50% v/v MPD and 0.1 M HEPES, pH 7.5, and one part protein solution containing 13 mg/ml enzyme LpxK, 4.3 mM AMP-PCP, 1 mM EDTA, 0.5% w/v DDM, 540 mM NaCl, 14% v/v glycerol, and 35 mM HEPES, pH 8.0, 20°C, 1 month, or by microseeding, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement using ADP-Mg2+ LpxK structure, PDB ID 4EHY, as the search model with all ligands removed, and AMP-PCP is subsequently added to the model. Purified LpxK in complex with ATP in a pre-catalytic binding state, mixing of seventeen parts of a reservoir solution consisting of 60% v/v MPD and 0.1 M HEPES, pH 7.5, and three parts protein solution containing 7.4 mg/ml enzyme LpxK, 10 mM ATP, 1 mM EDTA, 0.35% w/v DDM, 700 mM NaCl, 18.5% v/v glycerol, and 45 mM HEPES, pH 8.0, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement using LpxK structure, PDB ID 4EHX, as the search model, ATP is subsequently added to the model. Purified LpxK in complex with a chloride anion in an inhibitory conformation of the nucleotide-binding P-loop, mixing of three parts of a reservoir solution consisting of 40% v/v MPD and 0.1 M HEPES, pH 7.5, and one part protein solution containing 8.3 mg/ml LpxK, 4 mM methyl 2-acetamido-2-deoxy-beta-D-glucopyranoside, 0.35% w/v DDM, 625 mM NaCl, 17% v/v glycerol, and 45 mM HEPES, pH 8.0, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement using the apo enzyme LpxK structure, PDB ID 4EHX, as search model and spherical active site density refines well as a chloride ion |
