[NMR paper] Complete backbone and DENQ side chain NMR assignments in proteins from a single experiment: implications to structure-function studies.

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Complete backbone and DENQ side chain NMR assignments in proteins from a single experiment: implications to structure-function studies.

Related Articles Complete backbone and DENQ side chain NMR assignments in proteins from a single experiment: implications to structure-function studies.

J Struct Funct Genomics. 2014 Feb 18;

Authors: Reddy JG, Hosur RV

Abstract
Resonance assignment is the first and the most crucial step in all nuclear magnetic resonance (NMR) investigations on structure-function relationships in biological macromolecules. Often, the assignment exercise has to be repeated several times when specific interactions with ligands, substrates etc., have to be elucidated for understanding the functional mechanisms. While the protein backbone serves to provide a scaffold, the side chains interact directly with the ligands. Such investigations will be greatly facilitated, if there are rapid methods for obtaining exhaustive information with minimum of NMR experimentation. In this context, we present here a pulse sequence which exploits the recently introduced technique of parallel detection*of multiple nuclei, e.g. (1)H and (13)C, and results in two 3D-data sets simultaneously. These yield complete backbone resonance assignment ((1)H(N), (15)N, (13)CO, (1)H?/(13)C?, and (1)H?/(13)C? chemical shifts) and side chain assignment of D, E, N and Q residues. Such an exhaustive assignment has the potential of yielding accurate 3D structures using one or more of several algorithms which calculate structures of the molecules very reliably on the basis of NMR chemical shifts alone. The side chain assignments of D, E, N, and Q will be extremely valuable for interaction studies with different ligands; D*and E side chains are known to be involved in majority of catalytic activities. Utility of this experiment has been demonstrated with Ca(2+) bound M-crystallin, which contains largely D, E, N and Q residues at the metal binding sites.


PMID: 24535112 [PubMed - as supplied by publisher]



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