Protein conformation by EPR spectroscopy using gadolinium tags clicked to genetically encoded p-azido-L-phenylalanine

Chem Commun (Camb). 2015 Nov 14;51(88):15898-901. doi: 10.1039/c5cc07121f. Epub 2015 Sep 21.

Abstract

Quantitative cysteine-independent ligation of a Gd(3+) tag to genetically encoded p-azido-L-phenylalanine via Cu(I)-catalyzed click chemistry is shown to deliver an exceptionally powerful tool for Gd(3+)-Gd(3+) distance measurements by double electron-electron resonance (DEER) experiments, as the position of the Gd(3+) ion relative to the protein can be predicted with high accuracy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Azides / chemistry
  • Carrier Proteins / chemical synthesis*
  • Carrier Proteins / chemistry
  • Click Chemistry
  • Electron Spin Resonance Spectroscopy*
  • Escherichia coli / chemistry*
  • Gadolinium*
  • Glutamates / chemistry*
  • Mutagenesis, Site-Directed
  • Phenylalanine / analogs & derivatives*
  • Phenylalanine / chemistry
  • Phenylalanine / genetics
  • Protein Structure, Tertiary*
  • Spin Labels

Substances

  • Azides
  • Carrier Proteins
  • Glutamates
  • Spin Labels
  • 4-azidophenylalanine
  • Phenylalanine
  • Gadolinium