High-throughput PIXE as an essential quantitative assay for accurate metalloprotein structural analysis; development and application.
Summary of "High-throughput PIXE as an essential quantitative assay for accurate metalloprotein structural analysis; development and application."
Metalloproteins comprise over one third of proteins, with approximately half of all enzymes requiring metal to function. Accurate identification of these metal atoms and their environment is a prerequisite to understanding biological mechanism. Using ion beam analysis through particle induced X-ray emission (PIXE), we have quantitatively identified the metal atoms in 30 previously structurally characterized proteins using minimal sample volume and a high-throughput approach. Over half of these metals had been misidentified in the deposited structural models. Some of the PIXE detected metals not seen in the models were explainable as artifacts from promiscuous crystallization reagents. For others, using the correct metal improved the structural models. For multi-nuclear sites, anomalous diffraction signals enabled the positioning of the correct metals to reveal previously obscured biological information. PIXE is insensitive to chemical environment, but coupled with experimental diffraction data deposited alongside the structural model, it enables validation and potential remediation of metalloprotein models, improving structural and more importantly, mechanistic knowledge.
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This article was published in the following journal.
Name: Journal of the American Chemical Society
ISSN: 1520-5126
Pages:
Links
- PubMed Source: http://www.ncbi.nlm.nih.gov/pubmed/31794207
- DOI: http://dx.doi.org/10.1021/jacs.9b09186
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