2f47
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Xray crystal structure of T4 lysozyme mutant L20/R63A liganded to methylguanidinium
OverviewOverview
The binding of guanidinium ion has been shown to promote a large-scale, translation of a tandemly duplicated helix in an engineered mutant of T4, lysozyme. The guanidinium ion acts as a surrogate for the guanidino group, of an arginine side chain. Here we determine whether methyl- and, ethylguanidinium provide better mimics. The results show that addition of, the hydrophobic moieties to the ligand enhances the binding affinity, concomitant with reduction in ligand solubility. Crystallographic analysis, confirms that binding of the alternative ligands to the engineered site, still drives the large-scale conformational change. Thermal analysis and, NMR data show, in comparison to guanidinium, an increase in protein, stability and in ligand affinity. This is presumably due to the successive, increase in hydrophobicity in going from guanidinium to ethylguanidinium., A fluorescence-based optical method was developed to sense the, ligand-triggered helix translation in solution. The results are a first, step in the de novo design of a molecular switch that is not related to, the normal function of the protein.
About this StructureAbout this Structure
2F47 is a Single protein structure of sequence from Bacteriophage t4 with CL, MGX and BME as ligands. Active as Lysozyme, with EC number 3.2.1.17 Full crystallographic information is available from OCA.
ReferenceReference
Guanidinium derivatives bind preferentially and trigger long-distance conformational changes in an engineered T4 lysozyme., Yousef MS, Bischoff N, Dyer CM, Baase WA, Matthews BW, Protein Sci. 2006 Apr;15(4):853-61. PMID:16600969
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