TY  - JOUR
A1  - Mehr, Alexander
A1  - Henneberg, Fabian
A1  - Chari, Ashwin
A1  - Görlich, Dirk
A1  - Huyton, Trevor
T1  - The copper(II)‐binding tripeptide GHK, a valuable crystallization and phasing tag for macromolecular crystallography
Y1  - 2020-12-02
VL  - 76
IS  - 12
SP  - 1222
EP  - 1232
JF  - Acta Crystallographica Section D
DO  - 10.1107/S2059798320013741
DO  - 10.23689/fidgeo-4112
PB  - International Union of Crystallography
N2  - 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.
N2  - 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
UR  - http://resolver.sub.uni-goettingen.de/purl?gldocs-11858/8452
ER  -