Abstract:
:Simultaneous data acquisition in time-sharing (TS) multi-dimensional NMR experiments has been shown an effective means to reduce experimental time, and thus to accelerate structure determination of proteins. This has been accomplished by spin evolution time-sharing of the X and Y heteronuclei, such as (15)N and (13)C, in one of the time dimensions. In this work, we report a new 3D TS experiment, which allows simultaneous (13)C and (15)N spin labeling coherence in both t(1) and t(2) dimensions to give four NOESY spectra in a single 3D experiment. These spectra represent total NOE correlations between (1)H(N) and (1)H(C) resonances. This strategy of double time-sharing (2TS) results in an overall four-fold reduction in experimental time compared with its conventional counterpart. This 3D 2TS CN-CN-H HSQC-NOESY-HSQC pulse sequence also demonstrates improvements in water suppression, (15)N spectral resolution and sensitivity, which were developed based on 2D TS CN-H HSQC and 3D TS H-CN-H NOESY-HSQC experiments. Combining the 3D TS and the 3D 2TS NOESY experiments, NOE assignment ambiguities and errors are considerably reduced. These results will be useful for rapid protein structure determination to complement the effort of discerning the functions of diverse genomic proteins.
journal_name
J Biomol NMRjournal_title
Journal of biomolecular NMRauthors
Xia Y,Yee A,Arrowsmith CH,Gao Xdoi
10.1023/a:1025407905478subject
Has Abstractpub_date
2003-11-01 00:00:00pages
193-203issue
3eissn
0925-2738issn
1573-5001pii
5141155journal_volume
27pub_type
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