   2.7.1.159 | purified recombinant enzyme free or in complex with ATP analogue AMP-PCP and substrate inositol-1,3,4-trisphosphate in presence of Mg2+, 30 mg/ml protein in 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 10 mM DTT, mixed with a solution containing 20% PEG 3000, 0.1 M Tris-HCl, pH 7.5, and 0.1 M Ca(OAc)2, several months, crystals are seeded into sitting drops of 15 mg/ml protein, 25% PEG 3350, 5% PEG 400, 0.1 M Bis-Tris, pH 5.5, 10 mM DTT, and 10 mM L-cysteine, with or without ligands added as 10 mM AMP-PCP, 10 mM MgCl2, and 5 mM inositol-1,3,4-trisphosphate, direct X-ray diffraction analysis after this step or further seeding adding ADP and other ligands, overview, X-ray diffraction structure determination and analysis at 1.2-2.0 A resolution |
662834 |
