A 3D time-shared NOESY experiment designed to provide optimal resolution for accurate assignment of NMR distance restraints in large proteins

Subrata H. Mishra, Bradley J. Harden, Dominique P. Frueh

Research output: Contribution to journalArticle

Abstract

Structure determination of proteins by solution NMR has become an established method, but challenges increase steeply with the size of proteins. Notably, spectral crowding and signal overlap impair the analysis of cross-peaks in NOESY spectra that provide distance restraints for structural models. An optimal spectral resolution can alleviate overlap but requires prohibitively long experimental time with existing methods. Here we present a time-shared 3D experiment optimized for large proteins that provides 15N and 13C dispersed NOESY spectra in a single measurement. NOESY correlations appear in the detected dimension and hence benefit from the highest resolution achievable of all dimensions without increase in experimental time. By design, this experiment is inherently optimal for non-uniform sampling acquisition when compared to current alternatives. Thus, 15N and 13C dispersed NOESY spectra with ultra-high resolution in all dimensions were acquired in parallel within about 4 days instead of 80 days for a 52 kDa monomeric protein at a concentration of 350 μM.

Original languageEnglish (US)
Pages (from-to)265-274
Number of pages10
JournalJournal of Biomolecular NMR
Volume60
Issue number4
DOIs
StatePublished - Dec 15 2014

Keywords

  • Large proteins
  • NMR structure determination
  • Resolution
  • TROSY NOESY
  • Time-shared

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

Fingerprint Dive into the research topics of 'A 3D time-shared NOESY experiment designed to provide optimal resolution for accurate assignment of NMR distance restraints in large proteins'. Together they form a unique fingerprint.

  • Cite this