An NMR experiment for simultaneous TROSY-based detection of amide and methyl groups in large proteins

J Am Chem Soc. 2008 Aug 20;130(33):10872-3. doi: 10.1021/ja8036178. Epub 2008 Jul 24.

Abstract

A sensitive 2D NMR experiment for simultaneous time-shared TROSY-type detection of amide and methyl groups in high-molecular-weight proteins is described. The pulse scheme is designed to preserve the slowly decaying components of both 1H-15N and methyl 13CH3 spin systems in the course of indirect evolution and acquisition periods. The proposed methodology is applied to the study of substrate binding to {U-[15N,2H]; Ile-[13CH3]; Leu,Val-[13CH3/12CD3]}-labeled 82-kDa enzyme Malate Synthase G and is expected to accelerate NMR-based screening of large proteins labeled with 15N and selectively labeled with 13CH3 at methyl sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / analysis*
  • Macromolecular Substances / analysis
  • Magnetic Resonance Spectroscopy / methods*
  • Magnetic Resonance Spectroscopy / standards
  • Proteins / analysis*
  • Reference Standards

Substances

  • Amides
  • Macromolecular Substances
  • Proteins