Bhavesh, Neel S. ; Chatterjee, Amarnath ; Panchal, Sanjay C. ; Hosur, Ramakrishna V. (2003) Application of HN(C)N to rapid estimation of 1J(N-Cα) coupling constants correlated to ψ torsion angles in proteins: implication to structural genomics Biochemical and Biophysical Research Communications, 311 (3). pp. 678-684. ISSN 0006-291X
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Official URL: http://linkinghub.elsevier.com/retrieve/pii/S00062...
Related URL: http://dx.doi.org/10.1016/j.bbrc.2003.09.237
Abstract
We recently described a triple resonance experiment, HN(C)N, for sequential correlation of HN and 15N atoms in (15N, 13C) labeled proteins [J. Biomol. NMR. 20 (2001) 135]. Here, we describe an approach based on this experiment for estimation of one bond N-Cα J-couplings in medium size labeled proteins, which seem to show good correlations with ψ torsion angles along the protein backbone. The approach uses the ratio of the intensities of the sequential and diagonal peaks in the F2-F3 planes of the HN(C)N spectrum. The reliability of the approach has been demonstrated using a short peptide wherein the coupling constants have been measured by the present method and also independently from peak splittings in HSQC spectra. The two results agree within 10%. The applicability of the procedure to proteins has been demonstrated using doubly labeled FK506 binding protein (FKBP, molecular mass ~12 kDa). Coupling constant estimates have been obtained for 62 out of 100 non-proline residues and they show a correlation with ψ torsion angles, as has been reported before. This semi-quantitative application of HN(C)N extends the significance of the experiment especially, in the context of structural genomics, since the single experiment, not only provides a great enhancement in the speed of resonance assignment, but also provides quantitative structural information.
Item Type: | Article |
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Source: | Copyright of this article belongs to Elsevier Science. |
Keywords: | HN(C)N; J(N-C) Coupling; ψ Torsion Angles; FK506 Binding Protein; Structural Genomics |
ID Code: | 16797 |
Deposited On: | 15 Nov 2010 13:15 |
Last Modified: | 03 Jun 2011 08:55 |
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