The Magic Formula For Y-27632

An optimized crystallization condition at high ionic strength resulted in two differing crystal forms of comparable quality (Table 2 ?, data sets 1 and 2). The differing modification levels of cgAUS1-a1 and cgAUS1-a2 had no effect on the crystallization of these enzyme samples. Co-crystallization of active cgAUS1-a2 with 100?mM 1,4-resorcinol resulted in smaller crystals belonging to space group P3121 (Table 2 ?, data set 3). Table 2 Data collection and processing Only two crystals of latent cgAUS1-ln were obtained, which diffracted to 2.50?? resolution (helical data-collection mode; Table 2 ?, data set 4). The crystallization was not reproducible as nucleation bepotastine and crystal growth were difficult to control within the LLPS. The phase separation was even more pronounced in crystallization setups of recombinantly expressed cgAUS1-lr. Crystals diffracting to 2.93?? resolution and belonging to space group P1 were only obtained after several weeks (Table 2 ?, data set 5). It has recently been shown that the use Y-27632 research buy of the polyoxometalate hexatungstotellurate(VI) as a crystallization additive (Bijelic & Rompel, 2015 ?) improved the crystallization of tyrosinase from A. bisporus (Mauracher et al., 2014b ?) and that this compound was involved in crystal packing (Mauracher et al., 2014a ?). Similar observations have been reported for the co-crystallization of this compound with HEWL (Bijelic et al., 2015 ?). Substitution of 100?mM magnesium chloride by 1?mM hexatungstotellurate(VI) (Na6[TeW6O24]) drastically improved the nucleation and crystal growth of cgAUS1-lr within the LLPS. Crystals diffracting to 2.08?? resolution were obtained within several days (Table 2 ?, data set 6). However, the crystals were very unstable in the cryoprotectant solution and on addition of reservoir solution to the crystallization Olaparib drop, as they rapidly generated cracks and dissolved. However, one data set (Table 2 ?, data set 6) was obtained from an unscathed crystal by reducing the contact time with the cryoprotectant solution to a minimum (?1?s) before flash-cooling. Lowering the pH of the crystallization condition improved the stability of the crystals without any significant change in the unit-cell parameters. These crystals were soaked with hydrogen peroxide to obtain the oxy-form of the dinuclear copper centre (Table 2 ?, data set 7). The successful crystallization of recombinantly expressed cgAUS1 enables crystallization of the mutants of cgAUS1 reported recently (Kaintz et al., 2015 ?). Crystallographic data and X-ray data-collection statistics are summarized in Table 2 ?. The crystal structure of active cgAUS1-a1 was solved by molecular replacement using the crystal structure of catechol oxidase from I. batatas as a model (?47% sequence identity; PDB entry 1bt3). The crystal structure of latent cgAUS1-ln was solved by molecular replacement followed by automated model building. Refinement of the obtained models is in progress.