Abstract Development of efficient strategies and automation represent important milestones of progress in rapid structure determination efforts in proteomics research. In this context, we present here an efficient algorithm named as AUTOBA (Automatic Backbone Assignment) designed to automate the assignment protocol based on HN(C)N suite of experiments. Depending upon the spectral dispersion, the user can record 2D or 3D versions of the experiments for assignment. The algorithm uses as inputs: (i) protein primary sequence and (ii) peak-lists from user defined HN(C)N suite of experiments. In the end, one gets HN, 15N, Cα and C� assignments (in common BMRB format) for the individual residues along the polypeptide chain. The success of the algorithm has been demonstrated, not only with experimental spectra recorded on two small globular proteins: ubiquitin (76 aa) and M-crystallin (85 aa), but also with simulated spectra of 27 other proteins using assignment data from the BMRB.
Content Type Journal Article
Pages 1-13
DOI 10.1007/s10858-011-9518-0
Authors
Aditi Borkar, Department of Chemical Sciences, Tata Institute of Fundamental Research (TIFR), 1, Homi Bhabha Road, Colaba, Mumbai, 400005 India
Dinesh Kumar, Department of Chemical Sciences, Tata Institute of Fundamental Research (TIFR), 1, Homi Bhabha Road, Colaba, Mumbai, 400005 India
Ramakrishna V. Hosur, Department of Chemical Sciences, Tata Institute of Fundamental Research (TIFR), 1, Homi Bhabha Road, Colaba, Mumbai, 400005 India
5D 13C-detected experiments for backbone assignment of unstructured proteins with a very low signal dispersion
5D 13C-detected experiments for backbone assignment of unstructured proteins with a very low signal dispersion
Abstract Two novel 5D NMR experiments (CACONCACO, NCOCANCO) for backbone assignment of disordered proteins are presented. The pulse sequences exploit relaxation properties of the unstructured proteins and combine the advantages of 13C-direct detection, non-uniform sampling, and longitudinal relaxation optimization to maximize the achievable resolution and minimize the experimental time. The pulse sequences were successfully tested on the sample of partially disordered delta...
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Sparsely-sampled High-resolution 4-D Experiments for Efficient Backbone Resonance Assignment of Disordered Proteins
Sparsely-sampled High-resolution 4-D Experiments for Efficient Backbone Resonance Assignment of Disordered Proteins
Publication year: 2011
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 4 January 2011</br>
Jie, Wen , Jihui, Wu , Pei, Zhou</br>
Intrinsically disordered proteins (IDPs) play important roles in many critical cellular processes. Due to their limited chemical shift dispersion, IDPs often require four pairs of resonance connectivities (H?, C?, C? and CO) for establishing sequential backbone assignment. Because most conventional 4-D...
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01-05-2011 11:03 AM
Pseudo-4D triple resonance experiments to resolve HN overlap in the backbone assignment of unfolded proteins
Pseudo-4D triple resonance experiments to resolve HN overlap in the backbone assignment of unfolded proteins
Abstract The solution NMR resonance assignment of the protein backbone is most commonly carried out using triple resonance experiments that involve 15N and 1HN resonances. The assignment becomes problematic when there is resonance overlap of 15Nâ??1HN cross peaks. For such residues, one cannot unambiguously link the â??leftâ?? side of the NH root to the â??rightâ?? side, and the residues associated with such overlapping HN resonances remain often unassigned. Here we present a...
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[NMR paper] DNA bending and sequence-dependent backbone conformation NMR and computer experiments
DNA bending and sequence-dependent backbone conformation NMR and computer experiments.
Related Articles DNA bending and sequence-dependent backbone conformation NMR and computer experiments.
Eur J Biochem. 1999 Oct 1;265(1):35-53
Authors: Ojha RP, Dhingra MM, Sarma MH, Shibata M, Farrar M, Turner CJ, Sarma RH
Although DNA bending plays a crucial role in several biological processes, very little is known experimentally about the relationship between sugar phosphate conformation and sequence directed bending. In this paper, we determine the...
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11-18-2010 08:31 PM
Corrigendum to “BEST-HNN and 2D (HN)NH experiments for rapid backbone assignment in p
Corrigendum to “BEST-HNN and 2D (HN)NH experiments for rapid backbone assignment in proteins”
Publication year: 2010
Source: Journal of Magnetic Resonance, In Press, Corrected Proof, Available online 15 September 2010</br>
Dinesh, Kumar , Subhradip, Paul , Ramakrishna V., Hosur</br>
Source: Journal of Magnetic Resonance
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[NMR paper] Automation of protein 2D proton NMR assignment by means of fuzzy mathematics and grap
Automation of protein 2D proton NMR assignment by means of fuzzy mathematics and graph theory.
Related Articles Automation of protein 2D proton NMR assignment by means of fuzzy mathematics and graph theory.
J Chem Inf Comput Sci. 1993 Sep-Oct;33(5):668-82
Authors: Xu J, Straus SK, Sanctuary BC, Trimble L
The novel methodology for protein 2D NMR assignment presented in this paper is based upon protein spin coupling graph theory analysis, fuzzy graph pattern recognition, and tree searching. The method required to formalize the whole assignment...
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08-22-2010 03:01 AM
HA-detected experiments for the backbone assignment of intrinsically disordered prote
Abstract We propose a new alpha proton detection based approach for the sequential assignment of natively unfolded proteins. The proposed protocol superimposes on following features: HA-detection (1) enables assignment of natively unfolded proteins at any pH, i.e., it is not sensitive to rapid chemical exchange undergoing in natively unfolded proteins even at moderately high pH. (2) It allows straightforward assignment of proline-rich polypeptides without additional proline-customized experiments. (3) It offers more streamlined and less ambiguous assignment based on solely intraresidual...
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Suite of Six NMR Relaxation Dispersion Experiments to Study Multiple-Site Exchange in Proteins
http://pubs.acs.org/isubscribe/journals/jacsat/127/i44/figures/ja054550en00001.gif
Multiple-Site Exchange in Proteins Studied with a Suite of Six NMR Relaxation Dispersion Experiments: An Application to the Folding of a Fyn SH3 Domain Mutant
Dmitry M. Korzhnev, Philipp Neudecker, Anthony Mittermaier, Vladislav Yu. Orekhov, and Lewis E. Kay*
Contribution from the Departments of Medical Genetics, Biochemistry, and Chemistry, The University of Toronto, Toronto, Ontario M5S 1A8, Canada, and Swedish NMR Center at Göteborg University, Box 465, 405 30 Göteborg, Sweden
J. Am. Chem....
AUTOBA: Automation of backbone assignment from HN(C)N suite of experiments
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