Editing and diagonal peak suppression in three-dimensional HCCH protein NMR correlation experiments.
A novel three-dimensional (3D) HCCH NMR experiment is introduced. It involves 13C-13C COSY or TOCSY coherence transfer plus two independent editing steps according to the number of protons attached to the individual carbons before and after the 13C-13C homonuclear mixing. This double editing leads to simplification of HCCH protein side chain spectra that otherwise are prone to spectral overlap. Another interesting feature is amino acid selectivity, i.e. that the presence of certain correlations in a doubly edited HCCH subspectrum gives a clue as to assignment to a particular subgroup of amino acids or segments thereof. Finally, the selection of two different multiplicities in the two editing steps leads to diagonal peak suppression in the 1H-1H (3D spectrum recorded with two 1H and one 13C dimension) or the 13C-13C (3D spectrum recorded with one 1H and two 13C dimensions) two-dimensional projection. The new experiment is demonstrated using a 13C,15N-labeled protein sample, chymotrypsin inhibitor 2, at 500 MHz.
Meissner
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Department of Chemistry, Carlsberg Laboratory, Valby, Denmark.
Sørensen
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D000818:Animals; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008280:Magnetics; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010455:Peptides / Q000737:chemistry; D010940:Plant Proteins; D011506:Proteins / Q000737:chemistry*
D010455:Peptides; D010940:Plant Proteins; D011506:Proteins; C093336:chymotrypsin inhibitor 2