Abstract：
:Extensive resonance overlap exacerbates assignment of intrinsically disordered proteins (IDPs). This issue can be circumvented by utilizing (15)N, (13)C' and (1)H(N) spins, where the chemical shift dispersion is mainly dictated by the characteristics of consecutive amino acid residues. Especially (15)N and (13)C' spins offer superior chemical shift dispersion in comparison to (13)C(α) and (13)C(β) spins. However, HN-detected experiments suffer from exchange broadening of amide proton signals on IDPs especially under alkali conditions. To that end, we propose here two novel HA-detected experiments, (HCA)CON(CA)H and (HCA)NCO(CA)H and a new assignment protocol based on panoply of unidirectional HA-detected experiments that enable robust backbone assignment of IDPs also at high pH. The new approach was tested at pH 6.5 and pH 8.5 on cancer/testis antigen CT16, a 110-residue IDP, and virtually complete backbone assignment of CT16 was obtained by employing the novel HA-detected experiments together with the previously introduced iH(CA)NCO scheme. Remarkably, also those 10 N-terminal residues that remained unassigned in our earlier HN-detection based assignment approach even at pH 6.5 were now readily assigned. Moreover, theoretical calculations and experimental results suggest that overall sensitivity of the new experiments is also applicable to small or medium sized globular proteins that require alkaline conditions.
journal_name
J Biomol NMRjournal_title
Journal of biomolecular NMRauthors
Mäntylahti S,Hellman M,Permi Pdoi
10.1007/s10858-011-9470-zsubject
Has Abstractpub_date
2011-02-01 00:00:00pages
99-109issue
2eissn
0925-2738issn
1573-5001journal_volume
49pub_type
杂志文章abstract：:A comparison is made of the consequences of using time-averaged and conventional vicinal (3)J-coupling restraints in molecular dynamics refinement of an adenosine nucleoside model system. The target values for the restraints are derived from a 3-ns unrestrained molecular dynamics simulation. A comparison of the result...
journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/BF00175253
更新日期：1994-03-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-018-0197-y
更新日期：2018-06-01 00:00:00
abstract：:A simple spectroscopic filtering technique is presented that may aid the assignment of (13)C and (15)N resonances of methyl-containing amino-acids in solid-state magic-angle spinning (MAS) NMR. A filtering block that selects methyl resonances is introduced in two-dimensional (2D) (13)C-homonuclear and (15)N-(13)C hete...
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1023/a:1019854626147
更新日期：2002-07-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1023/a:1008318530946
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-018-0207-0
更新日期：2018-10-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-013-9705-2
更新日期：2013-03-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1023/a:1008202230140
更新日期：1998-04-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-011-9520-6
更新日期：2011-08-01 00:00:00
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journal_title：Journal of biomolecular NMR
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doi：10.1007/s10858-014-9856-9
更新日期：2014-11-01 00:00:00
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pub_type： 杂志文章
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更新日期：2019-11-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章,评审
doi：10.1007/s10858-009-9376-1
更新日期：2010-01-01 00:00:00
abstract：:The solvent-exposed regions of (U-13C)ascomycin when bound to its putative target protein, FKBP, have been identified based on the different proton longitudinal relaxation rates (R1 = 1/T1) measured in the absence and presence of the paramagnetic relaxation reagent, 4-hydroxy-2,2,6,6-tetramethyl-piperidinyl-1-oxy (HyT...
journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/BF02192797
更新日期：1992-01-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-015-9957-0
更新日期：2015-07-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-011-9567-4
更新日期：2011-11-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-016-0031-3
更新日期：2016-04-01 00:00:00
abstract：:During the past three years it has become possible to compute ab initio the 13C, 15N and 19F NMR chemical shifts of many sites in native proteins. Chemical shifts are beginning to become a useful supplement to more established methods of solution structure determination, and may find utility in solid-state analysis as...
journal_title：Journal of biomolecular NMR
pub_type： 杂志文章,评审
doi：10.1007/BF00211749
更新日期：1995-04-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-018-00223-3
更新日期：2019-02-01 00:00:00
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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
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doi：10.1023/B:JNMR.0000032614.79833.3d
更新日期：2004-07-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1023/B:JNMR.0000013701.16162.0c
更新日期：2004-03-01 00:00:00
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pub_type： 杂志文章
doi：10.1007/s10858-014-9845-z
更新日期：2014-08-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/BF01877224
更新日期：1991-07-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1023/a:1008285512845
更新日期：1998-08-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/BF00174609
更新日期：1993-09-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-020-00310-4
更新日期：2020-05-01 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：2003-11-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1007/s10858-018-0173-6
更新日期：2018-04-01 00:00:00
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journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
doi：10.1023/a:1008351128089
更新日期：2000-03-01 00:00:00
abstract：:With the completion of genome sequencing projects, there are a large number of proteins for which we have little or no functional information. Since protein function is closely related to three-dimensional conformation, structural proteomics is one avenue where the role of proteins with unknown function can be investi...
journal_title：Journal of biomolecular NMR
pub_type： 杂志文章
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更新日期：2006-06-01 00:00:00
