Publication year: 2012 Source: Journal of Magnetic Resonance, Available online 8 February 2012
Manoj Kumar*Rout, Pushpa*Mishra, Hanudatta S.*Atreya, Ramakrishna V.*Hosur
We present here an improvisation of HNN (Panchal, Bhavesh et al. 2001)called RD 3D HNCANfor backbone (HN, CA andN) assignment inboth folded and unfolded proteins. This is a reduced dimensionality experiment which employsCAchemical shifts to improve dispersion. Distinct positive and negative peak patternsof various triplet segments along the polypeptide chain observed in HNN are retained and these provide start and check points for the sequential walk. Because of co-incrementing of CA andN, peaks along one of the dimensions appear at sums and differences of the CAandN chemical shifts. This changes the backbone assignment protocol slightly and we present this in explicit detail. The performance of the experiment has been demonstrated using Ubiquitin and Plasmodium falciparum P2 proteins. The experiment is particularly valuable when two neighboring amino acid residues have nearly identical backboneN chemical shifts. Graphical abstract
Highlights
? 3D HNCANincorporates CA and N co-evolution in HNN. ? Provides HN, N and CA backbone assignments. ? Reduced dimensionality enhances spectral dispersion.
Efficient sequential assignments in proteins with reduced dimensionality 3D HN(CA)NH
Efficient sequential assignments in proteins with reduced dimensionality 3D HN(CA)NH
Abstract We present reduced dimensionality (RD) 3D HN(CA)NH for efficient sequential assignment in proteins. The experiment correlates the 15N and 1H chemical shift of a residue (â??iâ??) with those of its immediate N-terminal (i â?? 1) and C-terminal (i + 1) neighbors and provides four-dimensional chemical shift correlations rapidly with high resolution. An assignment strategy is presented which combines the correlations observed in this experiment with amino acid type information obtained from 3D ...
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A segmental labeling strategy for unambiguous determination of domainâ??domain interactions of large multi-domain proteins
A segmental labeling strategy for unambiguous determination of domainâ??domain interactions of large multi-domain proteins
Abstract NMR structural determination of large multi-domain proteins is a challenging task due to significant spectral overlap with a particular difficulty in unambiguous identification of domainâ??domain interactions. Segmental labeling is a NMR strategy that allows for isotopically labeling one domain and leaves the other domain unlabeled. This significantly simplifies spectral overlaps and allows for quick identification of domainâ??domain interaction. Here, a...
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07-08-2011 07:01 PM
Aliasing in reduced dimensionality NMR spectra: (3,2)D HNHA and (4,2)D HN(COCA)NH experiments as examples
Aliasing in reduced dimensionality NMR spectra: (3,2)D HNHA and (4,2)D HN(COCA)NH experiments as examples
Abstract Reduced dimensionality NMR spectra usually require very large spectral widths in the shared dimension. In this paper we show that aliasing can be introduced in reduced dimensionality NMR spectra either to decrease the acquisition time or increase the resolution of the experiments without losing information. The gains of introducing aliasing are illustrated with two examples, the (3,2)D HNHA and the (4,2)D HN(COCA)NH experiments. In both cases a reduction of the spectral...
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[NMR paper] Reduced-dimensionality NMR spectroscopy for high-throughput protein resonance assignm
Reduced-dimensionality NMR spectroscopy for high-throughput protein resonance assignment.
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Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):8009-14
Authors: Szyperski T, Yeh DC, Sukumaran DK, Moseley HN, Montelione GT
A suite of reduced-dimensionality (13)C,(15)N,(1)H-triple-resonance NMR experiments is presented for rapid and complete protein resonance assignment. Even when using short measurement times, these experiments allow one to retain...
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11-24-2010 08:49 PM
[NMR paper] New (15)N NMR exchange experiments for the unambiguous assignment of (1)H(N)/(15)N re
New (15)N NMR exchange experiments for the unambiguous assignment of (1)H(N)/(15)N resonances of proteins in complexes in slow chemical exchange with free form.
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J Magn Reson. 2000 Feb;142(2):276-9
Authors: Vialle-Printems C, van Heijenoort C, Guittet E
The potentialities of a 2D proton-detected heteronuclear exchange experiment to assign the nitrogen and amide proton...
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[NMR paper] Assignment of the 15N NMR spectra of reduced and oxidized Escherichia coli thioredoxi
Assignment of the 15N NMR spectra of reduced and oxidized Escherichia coli thioredoxin.
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FEBS Lett. 1991 Jun 24;284(2):178-83
Authors: Chandrasekhar K, Krause G, Holmgren A, Dyson HJ
As a necessary first step in the use of heteronuclear correlated spectra to obtain high resolution solution structures of the protein, assignment of the...
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08-21-2010 11:16 PM
[NMR paper] Assignment of active-site protons in the 1H-NMR spectrum of reduced human Cu/Zn super
Assignment of active-site protons in the 1H-NMR spectrum of reduced human Cu/Zn superoxide dismutase.
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Eur J Biochem. 1991 May 8;197(3):691-7
Authors: Bertini I, Capozzi F, Luchinat C, Piccioli M, Viezzoli MS
600-MHz 1H-NMR and nuclear Overhauser enhancement spectroscopy (NOESY)...
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hnCOcaNH and hncoCANH pulse sequences for rapid and unambiguous backbone assignment i
hnCOcaNH and hncoCANH pulse sequences for rapid and unambiguous backbone assignment in (13C, 15N) labeled proteins
Publication year: 2010
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 30 June 2010</br>
Dinesh, Kumar , Jithender G, Reddy , Ramakrishna V., Hosur</br>
Time-saving in data acquisition is a major thrust of NMR pulse sequence development in the context of structural proteomics research. The conventional HNCA and HN(CA)CO pulse sequences, routinely used for sequential backbone assignment, have the limitation that they cannot...
Reduced dimensionality 3D HNCANfor unambiguous HN, CA and N assignment in proteins
Linear Mode
