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Structural determination of a partially hemihedrally twinned actin crystal

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An orthorhombic actin crystal (space group P212121, unit-cell parameters a = 101.6, b = 103.0, c = 127.0 Å) was converted into a partially hemihedrally twinned tetragonal crystal (space group P43, unit-cell parameters a = b = 101.5, c = 104.2 Å) by induced condensation. This condensation (decrease in the c axis) was caused by the flash-freezing of the crystal, with 30% PEG 400 as a cryoprotectant, prior to data collection. Diffraction data for the twinned tetragonal crystal were collected at 100 K to 3.0 Å resolution (99.8% completeness with an Rsym of 8.1%) using synchrotron radiation. The hemihedral twinning of the data was observed by self-rotation function analysis and was determined to have a partial twin fraction of 0.376 from intensity statistics. The structure, with two actin molecules in the crystallographic asymmetric unit, was determined by molecular-replacement methods and refined to an R factor of 0.193. As a consequence of the crystal lattice transformation from the orthorhombic P212121 to the tetragonal P43 space group, actin–actin contacts were rearranged and an inter-actin dimer disulfide bond (Cys374) observed in the orthorhombic crystal form was broken in the tetragonal crystal form.

Keywords: actin; partial hemihedral twinning

Document Type: Research Article


Publication date: June 1, 2004


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