 | Acta Crystallographica Section F Acta Crystallographica Section F STRUCTURAL BIOLOGY COMMUNICATIONS |
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![[Figure 3]](cb5089fig3mag.jpg)
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Figure 3
Improved crystal structure of human Sirt2 in complex with the product analogue ADPR. (a) Superposition of the published X-ray structure of human Sirt2 in complex with ADPR (PDB entry 3zgv, chain A, yellow) with the improved X-ray structure of the Sirt2–ADPR complex (chain A, dark grey) shows that the structures are very similar (r.m.s.d. of 0.18 Å for all Cα atoms). (b) Close-up view of the active site of the improved Sirt2–ADPR complex. ADPR (overall B factor of 13.8 Å2) is shown as aquamarine sticks and the σ-weighted 2Fo − Fc electron-density map is contoured at 1.0σ. A σ-weighted Fo − Fc electron-density OMIT map of ADPR is shown in Supplementary Fig. S4. (c) Close-up view of the hinge loop of Sirt2. In the published X-ray structure of Sirt2 parts of this loop were not defined by the electron density. In the improved structure of Sirt2, the conformations of all amino acids that form this loop are well defined by the electron density. The σ-weighted 2Fo − Fc electron-density map is also contoured at 1.0σ. A σ-weighted Fo − Fc electron-density OMIT map of the hinge loop is shown in Supplementary Fig. S4. (d) Schematic representation of crystal hits for the complex of human Sirt256–356 and ADPR in a screen to optimize the initial crystallization conditions. Two different crystallization-drop compositions were used. In the presence of microseeds, Sirt256–356 crystallizes more rapidly and yields more crystals using diverse drop compositions, pH and PEG 10 000 concentrations. A red condition indicates crystals in both drops, while orange indicates crystal formation in only one. (e) Representative Sirt2–ADPR crystal after 5 d in a solution consisting of 15%(w/v) PEG 10 000 in 0.1 M bis-tris buffer pH 6.0. (f) UV image of (e). (g) Representative crystals of the Sirt2–ADPR complex obtained in the presence of microseeds in the solution mentioned in (e). (h) UV image of (g). |
![[Figure 3]](cb5089fig3.jpg)
| STRUCTURAL BIOLOGY COMMUNICATIONS |
ISSN: 2053-230X
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