Multidimensional 1H and 15N NMR investigation of glutamine-binding protein of Escherichia coli.
Related Articles Multidimensional 1H and 15N NMR investigation of glutamine-binding protein of Escherichia coli.
J Biomol NMR. 1992 Mar;2(2):149-60
Authors: Tjandra N, Simplaceanu V, Cottam PF, Ho C
Specific and uniform 15N labelings along with site-directed mutagenesis of glutamine-binding protein have been utilized to obtain assignments of the His156, Trp32 and Trp220 residues. These assignments have been made not only to further study the importance of these 3 amino acid residues in protein-ligand and protein-protein interactions associated with the active transport of L-glutamine across the cytoplasmic membrane of Escherichia coli, but also to serve as the starting points in the sequence-specific backbone assignment. The assignment of H epsilon 2 of His156 refines the earlier model where this particular proton forms an intermolecular hydrogen bond to the delta-carbonyl of L-glutamine, while assignments of both Trp32 and Trp220 show the variation in local structures which ensure the specificity in ligand binding and protein-protein interaction. Using 3D NOESY-HMQC NMR, amide connectivities can be traced along 8-9 amino acid residues at a time. This paper illustrates the usefulness of combining 15N isotopic labeling and multinuclear, multidimensional NMR techniques for a structural investigation of a protein with a molecular weight of 25,000.
PMID: 1422149 [PubMed - indexed for MEDLINE]
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PubMed