1.2.1.59 | purified His-tagged enzyme in complex with NAD+, microseeding method, mixing of 8.9 mg/ml protein in 25 mM HEPES, pH 7.35, 0.1 M NaCl, and 5 mM 2-mercaptoethanol, with crystallization solution containing 26-36% PEG 4000, 0.1 M MES, pH 6.5, the needle-like crystals are used for microseeding by mixing 0.002 ml of protein plus cofactor mixture, 500 nl water and 500 nl reservoir solution containing seed suspension in 28% PEG 4000, 0.1 M MES, pH 6.5, X-ray diffraction structure determination and analysis at 2.46 A resolution, molecular replacement and modelling |
741524 |