Abstract
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.
Original language | English (US) |
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Pages (from-to) | 151-158 |
Number of pages | 8 |
Journal | Journal of Structural Biology |
Volume | 192 |
Issue number | 2 |
DOIs | |
State | Published - Nov 2015 |
Externally published | Yes |
Keywords
- Electron microscopy
- Molecular labeling
- Unnatural amino acids
ASJC Scopus subject areas
- Structural Biology