A Genetically Encoded Spin Label for Electron Paramagnetic Resonance Distance Measurements
Journal of the American Chemical Society2014Vol. 136(4), pp. 1238–1241
Citations Over TimeTop 1% of 2014 papers
Abstract
We report the genetic encoding of a noncanonical, spin-labeled amino acid in Escherichia coli. This enables the intracellular biosynthesis of spin-labeled proteins and obviates the need for any chemical labeling step usually required for protein electron paramagnetic resonance (EPR) studies. The amino acid can be introduced at multiple, user-defined sites of a protein and is stable in E. coli even for prolonged expression times. It can report intramolecular distance distributions in proteins by double-electron electron resonance measurements. Moreover, the signal of spin-labeled protein can be selectively detected in cells. This provides elegant new perspectives for in-cell EPR studies of endogenous proteins.
Related Papers
- → Sub‐Micromolar Pulse Dipolar EPR Spectroscopy Reveals Increasing CuII‐labelling of Double‐Histidine Motifs with Lower Temperature(2019)87 cited
- → EPR Distance Measurements as a Tool to Characterize Protein‐DNA Interactions(2019)9 cited
- → CW EPR and DEER Methods to Determine BCL-2 Family Protein Structure and Interactions: Application of Site-Directed Spin Labeling to BAK Apoptotic Pores(2018)4 cited
- → Alignment Studies Employing the Rigid TOAC Spin Label Utilizing Electron Paramagnetic Resonance (EPR)(2010)
- High-field CW EPR with Gd(III) spin labels for structure studies of membrane proteins(2017)