Simultaneous Detection and Deconvolution of Congested NMR Spectra Containing Three Isotopically Labeled Species
- 31 May 2008
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 130 (25) , 7818-7819
- https://doi.org/10.1021/ja802701w
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 13Cα. By using the dual carbon label selective HSQC (DCLS-HSQC) pulse sequence and exploiting differences in 1J15N−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.Keywords
This publication has 20 references indexed in Scilit:
- Probing Chemical Shifts of Invisible States of Proteins with Relaxation Dispersion NMR Spectroscopy: How Well Can We Do?Journal of the American Chemical Society, 2008
- Asymmetric Methyl Group Labeling as a Probe of Membrane Protein Homo-oligomers by NMR SpectroscopyJournal of the American Chemical Society, 2008
- Isotopically Discriminated NMR Spectroscopy: A Tool for Investigating Complex Protein Interactions in VitroJournal of the American Chemical Society, 2007
- Quantitative dynamics and binding studies of the 20S proteasome by NMRNature, 2007
- Biomolecular NMR: a chaperone to drug discoveryCurrent Opinion in Chemical Biology, 2006
- NMR studies of protein structure and dynamicsPublished by Elsevier ,2004
- Studies of protein–ligand interactions by NMRBiochemical Society Transactions, 2003
- Identification by NMR Spectroscopy of Residues at Contact Surfaces in Large, Slowly Exchanging Macromolecular ComplexesJournal of the American Chemical Society, 1999
- Density Functional Calculations of Proton Chemical Shifts in Model PeptidesJournal of the American Chemical Society, 1997
- Measurement of three-bond nitrogen-carbon J couplings in proteins uniformly enriched in nitrogen-15 and carbon-13Journal of the American Chemical Society, 1993