Simultaneous detection and deconvolution of congested NMR spectra containing three isotopically labeled species

J Am Chem Soc. 2008 Jun 25;130(25):7818-9. doi: 10.1021/ja802701w. Epub 2008 May 31.

Abstract

We present a procedure for isolating subspectra corresponding to individual protein or peptide components in a ternary mixture or complex. Each of the three-component species is labeled differently: species A uniformly with 15N, species B uniformly with 15N and 13C, and species C uniformly with 15N but selectively with 13C' or 13Calpha. By using the dual carbon label selective HSQC (DCLS-HSQC) pulse sequence and exploiting differences in 1J 15N-13C coupling patterns to filter selected 15N resonances from detection during a constant time period, a subspectrum from each species can be generated from three spectra acquired from a single sample. Many important biological pathways involve dynamic interactions among members of multicomponent protein assemblies, and this approach offers a powerful way to monitor such processes.

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

  • Carbon Isotopes / chemistry
  • Hydrogen / chemistry
  • Isotope Labeling / methods*
  • Nitrogen Isotopes / chemistry
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Proteins / analysis*
  • Proteins / chemistry

Substances

  • Carbon Isotopes
  • Nitrogen Isotopes
  • Proteins
  • Hydrogen