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

MPS-Authors

Mehr,  A.
Department of Structural Dynamics, MPI for Biophysical Chemistry, Max Planck Society;

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Henneberg,  F.
Department of Structural Dynamics, MPI for Biophysical Chemistry, Max Planck Society;

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Chari,  A.
Research Group of Structural Biochemistry and Mechanisms, MPI for Biophysical Chemistry, Max Planck Society;

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Görlich,  D.
Department of Cellular Logistics, MPI for biophysical chemistry, Max Planck Society;

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Huyton,  T.
Department of Cellular Logistics, MPI for Biophysical Chemistry, Max Planck Society;

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Citation

Mehr, A., Henneberg, F., Chari, A., Görlich, D., & Huyton, T. (2020). The copper(II)-binding tripeptide GHK, a valuable crystallization and phasing tag for macromolecular crystallography. Acta Crystallographica D, 76(12), 1222-1232. doi:10.1107/S2059798320013741.


Cite as: http://hdl.handle.net/21.11116/0000-0008-0DDD-2
Abstract
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.