High-resolution methyl edited GFT NMR experiments for protein resonance assignments and structure determination

Jaipuria, Garima and Thakur, Anushikha and D'Silva, Patrick and Atreya, Hanudatta S (2010) High-resolution methyl edited GFT NMR experiments for protein resonance assignments and structure determination. In: Journal of Biomolecular NMR, 48 (3). pp. 137-145.

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Abstract

Three-dimensional (3D) structure determination of proteins is benefitted by long-range distance constraints comprising the methyl groups, which constitute the hydrophobic core of proteins. However, in methyl groups (of Ala, Ile, Leu, Met, Thr and Val) there is a significant overlap of C-13 and H-1 chemical shifts. Such overlap can be resolved using the recently proposed (3,2)D HCCH-COSY, a G-matrix Fourier transform (GFT) NMR based experiment, which facilitates editing of methyl groups into distinct spectral regions by combining their C-13 chemical shifts with that of the neighboring, directly attached, C-13 nucleus. Using this principle, we present three GFT experiments: (a) (4,3)D NOESY-HCCH, (b) (4,3)D H-1-TOCSY-HCCH and (c) (4,3)D C-13-TOCSY-HCCH. These experiments provide unique 4D spectral information rapidly with high sensitivity and resolution for side-chain resonance assignments and NOE analysis of methyl groups. This is exemplified by (4,3)D NOESY-HCCH data acquired for 17.9 kDa non-deuterated cytosolic human J-protein co-chaperone, which provided crucial long-range distance constraints for its 3D structure determination.

Item Type:	Journal Article
Publication:	Journal of Biomolecular NMR
Publisher:	Springer
Additional Information:	Copyright of this article belongs to Springer .
Keywords:	NOESY; TOCSY; GFT NMR; Methyl-edited NOESY; Protein structure
Department/Centre:	Division of Biological Sciences > Biochemistry Division of Chemical Sciences > NMR Research Centre (Formerly Sophisticated Instruments Facility) Division of Chemical Sciences > Solid State & Structural Chemistry Unit
Date Deposited:	13 Nov 2010 10:26
Last Modified:	23 Oct 2018 14:49
URI:	http://eprints.iisc.ac.in/id/eprint/33659

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