1te7

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Solution NMR Structure of Protein yqfB from Escherichia coli. Northeast Structural Genomics Consortium Target ET99

Overview

A protocol for high-quality structure determination based on G-matrix, Fourier transform (GFT) NMR is presented. Five through-bond chemical shift, correlation experiments providing 4D and 5D spectral information at high, digital resolution are performed for resonance assignment. These are, combined with a newly implemented (4,3)D GFT NOESY experiment which, encodes information of 4D 15N/15N-, 13C(alipahtic)/15N-, and, 13C(aliphatic)/13C(aliphatic)-resolved [1H,1H]-NOESY in two subspectra, each containing one component of chemical shift doublets arising from 4D, --> 3D projection at omega1:Omega(1H) +/- Omega(X) [X =, 15N,13C(aliphatic)]. The peaks located at the centers of the doublets are, obtained from simultaneous 3D 15N/13C(aliphatic)/13C(aromatic)-resolved, [1H,1H]-NOESY, wherein NOEs detected on aromatic protons are also, obtained. The protocol was applied for determining a high-quality, structure of the 14 kDa Northeast Structural Genomics consortium target, protein, YqfB (PDB ID ). Through-bond correlation and NOESY spectra were, acquired, respectively, in 16.9 and 39 h (30 h for shift doublets, 9 h for, central peaks) on a 600 MHz spectrometer equipped with a cryogenic probe., The rapidly collected highly resolved 4D NOESY information allows one to, assign the majority of NOEs directly from chemical shifts, which yields, accurate initial structures "within" approximately 2 angstroms of the, final structure. Information theoretical "QUEEN" analysis of initial, distance limit constraint networks revealed that, in contrast to, structure-based protocols, such NOE assignment is not biased toward, identifying additional constraints that tend to be redundant with respect, to the available constraint network. The protocol enables rapid NMR data, collection for robust high-quality structure determination of proteins up, to approximately 20-25 kDa in high-throughput.

About this Structure

1TE7 is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.

Reference

G-matrix Fourier transform NOESY-based protocol for high-quality protein structure determination., Shen Y, Atreya HS, Liu G, Szyperski T, J Am Chem Soc. 2005 Jun 29;127(25):9085-99. PMID:15969587

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