ISSN:
1573-5001
Keywords:
Heteronuclear NMR
;
Deuterium labeling
;
Cross-polarization
;
Triple-resonance experiment
;
Pulse scheme
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
Summary A new four-dimensional pulse scheme is described for the main-chain assignment of proteins by means of the J connectivity of the amide proton and nitrogen resonances of adjacent residues. Since the new experiment, 4D CP-HN(COCA)NH, involves heteronuclear cross-polarization for magnetization transfer from 13C=O to 15N via 13Cα, a relatively strong WALTZ-16 decoupling rf field is applied to 13Cα during magnetization transfer. Consequently, 13Cα is effectively decoupled from its attached 2H in the case of deuterated proteins, in the absence of a decoupling rf field for 2H. This efficiently improves the sensitivity of the experiment through 13C line narrowing. The experiment was performed on a randomly 60% deuterated protein, and the sensitivity of the final 4D spectrum was found to be excellent.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00211761
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