Fig. 7: Intracellular crystal growth and data collection allow solving protein structures with genuine cofactors. | Nature Communications

Fig. 7: Intracellular crystal growth and data collection allow solving protein structures with genuine cofactors.

From: A streamlined approach to structure elucidation using in cellulo crystallized recombinant proteins, InCellCryst

Fig. 7

a A MicroMesh mount loaded with High Five cells containing IMPDH ori crystals. Both the helical line scan and resulting dozor heat map generated at the P14 beamline (PETRA III, DESY, Hamburg) are shown. b A CrystalDirectTM plate for RT diffraction data collection (asterisk) is mounted on the Arinax MD3 diffractometer in an upright position (upper panel). Below, the loaded CrystalDirectTM plate is shown with HighFive cells carrying HEX-1 cyto crystals (close-up, right panel, arrow) adherent to the bottom foil. Next to each cell-containing well is a water-filled reservoir to keep humidity high and prevent cells from drying. c Cartoon representation of the HEX-1 ori structure. d IMPDH ori structure with natural ligands (ATP and GDP) in the Bateman domain and phosphates bound in the IMP binding site of the catalytic domain (spheres). Bateman domain and Finger domain in full colors, catalytic domains in light colors. e GDP within the canonical binding site 2 of the IMPDH ori Bateman domain. The omit map calculated with simulated annealing is shown at 3.0 sigma. f ATP within the canonical binding site 1 of the IMPDH ori Bateman domain. For clarity, phosphate moieties in the alternate conformation are shown in 50% transparency. The ligand omit map is shown at 3.0 sigma.

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