Line narrowing in methyl-TROSY using zero-quantum 1H-13C NMR spectroscopy.
An enhanced sensitivity zero-quantum correlation experiment is proposed for recording (1)H-(13)C correlations of methyl groups in highly deuterated, methyl protonated large proteins. The zero-quantum spectra benefit from TROSY-effects in which both intra- and inter-methyl dipolar relaxation interactions are minimized. Applications to malate synthase G at 5 degrees C (81 kDa single polypeptide chain enzyme, correlation time of 118 ns) and lysine decarboxylase at 45 degrees C (810 kDa decameric enzyme) are presented showing significant improvements in resolution relative to corresponding HMQC data sets, with only slight decreases (approximately 10%) in sensitivity.
Tugarinov
Vitali
V
Protein Engineering Network Centers of Excellence and Department of Medical Genetics, The University of Toronto, Toronto, Ontario, Canada, M5S 1A8.
Sprangers
Remco
R
Kay
Lewis E
LE
D002247:Carbon Isotopes; D002262:Carboxy-Lyases / Q000737:chemistry*; D003903:Deuterium; D008292:Malate Synthase / Q000737:chemistry*; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011522:Protons
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011522:Protons; D003903:Deuterium; D008292:Malate Synthase; D002262:Carboxy-Lyases; C021520:lysine decarboxylase