Site-specific labeling of proteins for electron microscopy

Details

• Alternative Title:
  J Struct Biol
• Personal Author:
  Dambacher, Corey M. ; Lander, Gabriel C.
• Description:
  Electron microscopy is commonly employed to determine the subunit organization of large macromolecular assemblies. However, the field lacks a robust molecular labeling methodology for unambiguous identification of constituent subunits. We present a strategy that exploits the unique properties of an unnatural amino acid in order to enable site-specific attachment of a single, readily identifiable protein label at any solvent-exposed position on the macromolecular surface. Using this method, we show clear labeling of a subunit within the 26S proteasome lid subcomplex that has not been amenable to labeling by traditional approaches.
• Subjects:
  Amino Acids Article Click Chemistry Electron Microscopy Escherichia Coli Imaging, Three-Dimensional Macromolecular Substances Microscopy, Electron Models, Molecular Molecular Labeling Proteasome Endopeptidase Complex Proteins Staining And Labeling Unnatural Amino Acids

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• Source:
  J Struct Biol. 192(2):151-158
• Pubmed ID:
  26409249
• Pubmed Central ID:
  PMC4633364
• Document Type:
  Journal Article
• Funding:
  DP2 EB020402/EB/NIBIB NIH HHS/United States ; DP2 EB020402-01/DP/NCCDPHP CDC HHS/United States
• Volume:
  192
• Issue:
  2
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:2fc9c7dd5494ab726826132da2a6d67332d9830fc12ba81b1e59cc2e5853d31a
• Download URL:
  https://stacks.cdc.gov/view/cdc/37796/cdc_37796_DS1.pdf
• File Type:
  [PDF - 1.03 MB ]