Journal
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
Volume 75, Issue -, Pages 764-771Publisher
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S2059798319009926
Keywords
metal quantification; quantitative X-ray anomalous dispersion; R2-like ligand-binding oxidase; Q-XAD; X-ray crystallography
Funding
- Vetenskapsradet [2016-03770, 2017-04018]
- European Research Council [HIGH-GEAR 724394]
- Knut och Alice Wallenbergs Stiftelse [2012.0233, 2017.0275]
- European Community Seventh Framework Programme (FP7/2007-2013) for BioStruct-X [283570]
- Swedish Research Council [2016-03770, 2017-04018] Funding Source: Swedish Research Council
Ask authors/readers for more resources
Here, a method is described which exploits X-ray anomalous dispersion (XAD) to quantify mixtures of metal ions in the binding sites of proteins and can be applied to metalloprotein crystals of average quality. This method has successfully been used to study site-specific metal binding in a protein from the R2-like ligand-binding oxidase family which assembles a heterodinuclear Mn/Fe cofactor. While previously only the relative contents of Fe and Mn in each metal-binding site have been assessed, here it is shown that the method can be extended to quantify the relative occupancies of at least three different transition metals, enabling complex competition experiments. The number of different metal ions that can be quantified is only limited by the number of high-quality anomalous data sets that can be obtained from one crystal, as one data set has to be collected for each transition-metal ion that is present (or is suspected to be present) in the protein, ideally at the absorption edge of each metal. A detailed description of the method, Q-XAD, is provided.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available