Rapid NMR assignments of proteins using optimized combinatorial selective unlabeling.
Rapid NMR assignments of proteins using optimized combinatorial selective unlabeling.
Chembiochem. 2015 Dec 10;
Authors: Atreya HS, Dubey A, Jaipuria G, Kadumuri RV, Vadrevu R
Abstract
A new approach for rapid resonance assignments in proteins based on amino acid selective unlabeling is presented. The method involves choosing a set of multiple amino acid types for selective unlabeling and identifying specific tri-peptides surrounding the labeled residues using two-dimensional (2D) NMR spectra in a combinatorial manner. It is shown that a 2D [15N-1H] HSQC spectrum along with two 2D spectra can yield ~50% assignments directly. The methodology is applicable to deuterated proteins and was applied to two systems: an intrinsically disordered protein (12 kDa) and 29 kDa (268 residues) ?-subunit of Escherichia coli tryptophan synthase, presenting a challenging case with spectral overlaps and missing peaks. Taken together, the method can augment conventional approaches and/or will be useful for applications such as identifying active-site residues, residues involved in ligand binding or protein-protein interactions even prior to complete resonance assignments.
PMID: 26662553 [PubMed - as supplied by publisher]
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