Abstract:
:The NMR spectra of the complex between the DNA-binding domain of the Dead ringer protein (DRI-DBD, Gly262-Gly398) and its DNA binding site (DRI-DBD:DNA, 26 kDa) have been optimized by biochemical and spectroscopic means. First, we demonstrate the utility of a modified 2D [F1,F2] 13C-filtered NOESY experiment that employs a 1J(HC) versus chemical shift optimized adiabatic 13C inversion pulse [Zwahlen, C. et al. (1997) J. Am. Chem. Soc., 119, 6711-6721]. The new sequence is shown to be more sensitive than previously published pulse schemes (up to 40% in favorable cases) and its utility is demonstrated using two protein-DNA complexes. Second, we demonstrate that the targeted replacement of an interfacial aromatic residue in the DRI-DBD:DNA complex substantially reduces line broadening within its NMR spectra. The spectral changes are dramatic, salvaging a protein-DNA complex that was originally ill suited for structural analysis by NMR. This biochemical approach is not a general method, but may prove useful in the spectral optimization of other protein complexes that suffer from interfacial line broadening caused by dynamic changes in proximal aromatic rings.
journal_name
J Biomol NMRjournal_title
Journal of biomolecular NMRauthors
Iwahara J,Wojciak JM,Clubb RTdoi
10.1023/a:1011296112710subject
Has Abstractpub_date
2001-03-01 00:00:00pages
231-41issue
3eissn
0925-2738issn
1573-5001journal_volume
19pub_type
杂志文章abstract::Recent efforts to reduce the measurement time for multidimensional NMR experiments have fostered the development of a variety of new procedures for sampling and data processing. We recently described concentric ring sampling for 3-D NMR experiments, which is superior to radial sampling as input for processing by a mul...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-008-9275-x
更新日期:2008-12-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00227466
更新日期:1995-01-01 00:00:00
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journal_title:Journal of biomolecular NMR
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00198369
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00404274
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00202041
更新日期:1996-05-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2011-02-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:1992-05-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-008-9224-8
更新日期:2008-03-01 00:00:00
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journal_title:Journal of biomolecular NMR
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journal_title:Journal of biomolecular NMR
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2015-12-01 00:00:00
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journal_title:Journal of biomolecular NMR
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2004-05-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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journal_title:Journal of biomolecular NMR
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2017-09-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00179343
更新日期:1994-05-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2005-02-01 00:00:00