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Mehr, Alexander [Verfasser:in]; Henneberg, Fabian [Verfasser:in]; Chari, Ashwin [Verfasser:in]; Görlich, Dirk [Verfasser:in]; Huyton, Trevor [Verfasser:in]; 1Department of Structural Dynamics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany [Verfasser:in]; 2Department of Cellular Logistics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany [Verfasser:in]

The copper(II)‐binding tripeptide GHK, a valuable crystallization and phasing tag for macromolecular crystallography

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  • Medientyp: E-Artikel
  • Titel: The copper(II)‐binding tripeptide GHK, a valuable crystallization and phasing tag for macromolecular crystallography
  • Beteiligte: Mehr, Alexander [Verfasser:in]; Henneberg, Fabian [Verfasser:in]; Chari, Ashwin [Verfasser:in]; Görlich, Dirk [Verfasser:in]; Huyton, Trevor [Verfasser:in]; 1Department of Structural Dynamics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany [Verfasser:in]; 2Department of Cellular Logistics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany [Verfasser:in]
  • Erschienen: International Union of Crystallography; 5 Abbey Square, Chester, Cheshire CH1 2HU, England, 2020-12-02
  • Sprache: Englisch
  • DOI: https://doi.org/10.1107/S2059798320013741
  • Schlagwörter: GHK ; phasing ; crystallization ; SAD
  • Entstehung:
  • Anmerkungen: Diese Datenquelle enthält auch Bestandsnachweise, die nicht zu einem Volltext führen.
  • Beschreibung: The growth of diffraction‐quality crystals and experimental phasing remain two of the main bottlenecks in protein crystallography. Here, the high‐affinity copper(II)‐binding tripeptide GHK was fused to the N‐terminus of a GFP variant and an MBP‐FG peptide fusion. The GHK tag promoted crystallization, with various residues (His, Asp, His/Pro) from symmetry molecules completing the copper(II) square‐pyramidal coordination sphere. Rapid structure determination by copper SAD phasing could be achieved, even at a very low Bijvoet ratio or after significant radiation damage. When collecting highly redundant data at a wavelength close to the copper absorption edge, residual S‐atom positions could also be located in log‐likelihood‐gradient maps and used to improve the phases. The GHK copper SAD method provides a convenient way of both crystallizing and phasing macromolecular structures, and will complement the current trend towards native sulfur SAD and MR‐SAD phasing. ; A novel three‐residue tag containing the residues GHK that can be used to promote crystallization and in SAD phasing experiments using its tightly bound copper ion is described. image
  • Zugangsstatus: Freier Zugang