General method for suppression of diagonal peaks in heteronuclear-edited NOESY spectroscopy.
A novel approach is proposed for suppression of diagonal peaks in 15N- and 13C-edited NOESY spectroscopy based on subtracting a spectrum with only diagonal peaks from the conventional NOESY spectrum. This method can be applied to most heteronuclear-edited NOESY experiments. It is far more sensitive than the TROSY-based approach for biomolecules with little TROSY effect, and nearly complete suppression of diagonal peaks can be achieved. The method has been demonstrated on samples of 15N-labeled calmodulin (17 kDa), 13C-labeled DdCAD-1 (24 kDa), and 13C-labeled hemoglobin (65 kDa), showing that cross-peaks very close to diagonals can be assigned reliably based on the difference spectra.
Wu
Jihui
J
Department of Biological Sciences and Department of Chemistry, National University of Singapore, 14 Science Drive 4, Singapore 117543.
Fan
Jing-song
JS
Pascal
Steven M
SM
Yang
Daiwen
D
D002147:Calmodulin / Q000737:chemistry; D002247:Carbon Isotopes; D006454:Hemoglobins / Q000737:chemistry; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D025801:Ubiquitin / Q000737:chemistry
D002147:Calmodulin; D002247:Carbon Isotopes; D006454:Hemoglobins; D009587:Nitrogen Isotopes; D011506:Proteins; D025801:Ubiquitin