2.7.4.21 | N-terminally truncated enzyme mutant, residues 41-366, hanging drop vapor diffusion against a well buffer of 12% w/v PEG 3350, 20 mM MgCl2, 0.1 M HEPES, pH 7.0, 0,1 mM AMP-PNP, and 2 mM CdCl2 at 4°C, 3 days, 4°C, X-ray diffraction structure determination and analysis |
738042 |
2.7.4.21 | purified recombinant MBP-tagged EhIP6KA residues 32-270, several crystal complexes of the IP6K including those that contain either large (Ins(1,3,4,5,6)P5 /InsP6) or small (Ins(1,4,5)P3) substrates, hanging drop vapor diffusion, 60 mg/ml protein in solution is mixed with reservoir solution containing 12% w/v PEG 3350, 50 mM NaH2PO4, pH 5.5 at 18°C, or by hanging drop vapor diffusion for one week against a well buffer containing 8% w/v PEG 3350, 100 mM Na3citrate, pH 5.2 at 25°C, followed by dilution microseeding for two weeks with a well buffer of 8% w/v PEG 3350, 100 mM Na3citrate, pH 5.2, and 8% ethylene glycol at 25°C, soaking of crystals in 22% w/v PEG 3350, 10 mM MgCl2, 10 mM ATP, 0.1 M sodium acetate, pH 5.2, and 20 mM InsP6, 10 mM Ins(1,3,4,5,6)P5 or 10 mM Ins(1,4,5)P3 for 3 days, X-ray diffraction structure determination and analysis, molecular replacement using MBP (1ez9) and IP3K (1w2c) as search models |
739144 |