Extensive 1H NMR resonance assignment of proteins using natural abundance gradient‐enhanced 13C−1H correlation spectroscopy

[摘要]

The reliability and completeness of ¹H NMR resonance assignment can be improved by the use of ¹³C−¹H HSQC correlation spectra on unlabelled protein samples using pulsed field gradients. This technique is illustrated on a 5.2 mM sample of the 79 residue Desulfovibrio vulgaris ferrocytochrome c ₅₅₃. Protons attached to the same carbon can be unambiguously paired in a HSQC spectrum. Contrary to ¹H, most amino acids exhibit characteristic ¹³C chemical shift ranges, which can be used for ¹³C assignment. This technique is especially useful for long side chain residues, such as Gln, Glu, Lys, Arg.

[学科分类] 生物化学/生物物理

[关键词] Nuclear magnetic resonance;Cytochrome;Chemical shift;Secondary structure;Carbon-13;Resonance assignment;NMR;nuclear magnetic resonance;NOESY;homonuclear nuclear Overhauser enhancement specroscopy;HOHAHA;homonuclear Hartman-Hahn spectroscopy;HMQC;1H-detected heteronuclear multiple-quantum coherence spectroscopy;HSQC;1H-detected heteronuclear single-quantum coherence spectroscopy;PFG;pulsed B0 field gradient;INEPT;insensitive nuclei enhanced by polarization transfer