An NMR experiment for simultaneous TROSY-based detection of amide and methyl groups in large proteins.
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.
Guo
Chenyun
C
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Zhang
Daoning
D
Tugarinov
Vitali
V
D000577:Amides / Q000032:analysis*; D046911:Macromolecular Substances / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy / Q000379:methods* / Q000592:standards; D011506:Proteins / Q000032:analysis*; D012015:Reference Standards
D000577:Amides; D046911:Macromolecular Substances; D011506:Proteins