   3.4.22.B70 | SENP1 crystallization is performed at 20°C using a sitting drop vapour-diffusion method. Single diamond-shaped crystals are grown after 2 days from equal volumes of protein solution (20 mg/ml in 20 mM Tris/HCl, pH 8.0, and 50 mM NaCl) and reservoir solution containing 100 mM CoCl2, 0.1 M Mes, pH 6.5, and 1.8 M (NH4)2SO4. The structure of SENP1 is determined to 2.45 A. NaBH4 is used to trap a stable thiohemiacetal transition-state analogue between Cys602 of SENP1 and Gly92 of SUMO-2, determination of the structure of this complex to 3.2 A. Crystallization and structure determination of the SENP1-SUMO-2 complex |
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