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

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.

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 Creators:
Mehr, A.1, Author
Henneberg, F.1, Author              
Chari, A.2, Author              
Görlich, D.3, Author              
Huyton, T.4, Author              
Affiliations:
1Department of Structural Dynamics, MPI for Biophysical Chemistry, Max Planck Society, ou_2205645              
2Research Group of Structural Biochemistry and Mechanisms, MPI for Biophysical Chemistry, Max Planck Society, ou_3265855              
3Department of Cellular Logistics, MPI for biophysical chemistry, Max Planck Society, ou_578574              
4Department of Cellular Logistics, MPI for Biophysical Chemistry, Max Planck Society, ou_578574              

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Free keywords: phasing; crystallization; GHK; SAD
 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.

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Language(s): eng - English
 Dates: 2020-12
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1107/S2059798320013741
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Title: Acta Crystallographica D
Source Genre: Journal
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Pages: - Volume / Issue: 76 (12) Sequence Number: - Start / End Page: 1222 - 1232 Identifier: -