Chemical shift based editing of CH3 groups in fractionally $^{13}C$-labelled proteins using GFT (3,2)D CT-HCCH-COSY: stereospecific assignments of $CH_3$ groups of Val and Leu residues

Barnwal, Ravi Pratap and Atreya, Hanudatta S and Chary, Kandala VR (2008) Chemical shift based editing of CH3 groups in fractionally $^{13}C$-labelled proteins using GFT (3,2)D CT-HCCH-COSY: stereospecific assignments of $CH_3$ groups of Val and Leu residues. In: Journal Of Biomolecular NMR, 42 (2). pp. 149-154.

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Abstract

We propose a (3,2)D CT-HCCH-COSY experiment to rapidly collect the data and provide significant dispersion in the spectral region containing $^13$C-$1^H$ cross peaks of $CH_3$ groups belonging to Ala, Ile, Leu, Met, Thr and Val residues. This enables one to carry out chemical shift based editing and grouping of all the $^{13}C$-$^1H$ cross peaks of $CH_3$ groups belonging to Ala, Ile, Leu, Met, Thr and Val residues in fractionally (10%) $^{13}C$-labelled proteins, which in turn aids in the sequence-specific resonance assignments in general and side-chain resonance assignments in particular, in any given protein. Further, we demonstrate the utility of this experiment for stereospecific assignments of the pro-R and pro-S methyl groups belonging to the Leu and Val residues in fractionally (10%) $^{13}C-labelled proteins. The proposed experiment opens up a wide range of applications in resonance assignment strategies and structure determination of proteins.

Item Type:	Journal Article
Additional Information:	Copyright for this article belongs to Springer
Keywords:	isotope labeling; NMR; automated assignments; sequence specific resonance assignments; stereo-specific resonance assignments
Department/Centre:	Division of Chemical Sciences
Date Deposited:	23 Dec 2008 07:41
Last Modified:	19 Sep 2010 04:51
URI:	http://eprints.iisc.ernet.in/id/eprint/16407

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