Cysteine-specific Cu2+ chelating tags used as paramagnetic probes in double electron electron resonance

J Phys Chem B. 2015 Feb 19;119(7):2839-43. doi: 10.1021/jp5103143. Epub 2015 Jan 30.

Abstract

Double electron electron resonance (DEER) is an attractive technique that is utilized for gaining insight into protein structure and dynamics via nanometer-scale distance measurements. The most commonly used paramagnetic tag in these measurements is a nitroxide spin label, R1. Here, we present the application of two types of high-affinity Cu(2+) chelating tags, based on the EDTA and cyclen metal-binding motifs as alternative X-band DEER probes, using the B1 immunoglobulin-binding domain of protein G (GB1) as a model system. Both types of tags have been incorporated into a variety of protein secondary structure environments and exhibit high spectral sensitivity. In particular, the cyclen-based tag displays distance distributions with comparable distribution widths and most probable distances within 1-3 Å when compared to homologous R1 distributions. The results display the viability of the cyclen tag as an alternative to the R1 side chain for X-band DEER distance measurements in proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cations*
  • Chelating Agents*
  • Copper*
  • Cyclams
  • Cysteine*
  • Edetic Acid
  • Heterocyclic Compounds
  • Magnetic Resonance Spectroscopy / methods*
  • Protein Structure, Secondary
  • Proteins / chemistry
  • Spin Labels

Substances

  • Cations
  • Chelating Agents
  • Cyclams
  • Heterocyclic Compounds
  • Proteins
  • Spin Labels
  • Copper
  • cyclen
  • Edetic Acid
  • Cysteine