   2.7.1.127 | purified recombinant unlabeled or selennomethionine-labeled mutant catalytic domain residues 185-459, 10 mg/ml protein in 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 10 mM DTT, crystallization solution contains 2.2 M ammonium sulfate, 0.2 M sodium formate, 10 mM DTT, 10 mM ADP, 10 mM 1D-myo-inositol 1,3,4,5-tetrakisphosphate, and 10 mM MgCl2, cryoprotection of crystals by successive transfer into 5-15% glycerol in mother liquor, X-ray diffraction structure determination and analysis at 2.2 A resolution |
| 2.7.1.127 | purified recombinant unlabeled or selennomethionine-labeled wild-type and mutant catalytic domain residues 187-461 free or complexed to ATP, substrate, and product, automated sitting drop vapour diffusion method, 17°C, 0.001 ml protein solution over reservoir solution containing 0.83-0.85 M tri-sodium citrate, 0.1 M Tris-HCl, pH 8.0, and 0.1 M NaCl, heavy atom derivative, substrates, or products complex formation by addition and removal of 0.001 ml of 2 M Li2SO4 and 100 mM Tris-HCl, pH 8.0, 4°C, containing the relevant compound, i.e. 3 mM ATP, 3 mM MnCl2, or 5 mM 1D-myo-inositol 1,3,4,5-tetrakisphosphate, for 7times, and final soaking for 2.5 h, X-ray diffraction structure determination and analysis at 1.8-1.9 A resolution |
