Using Singular Value Decomposition to Characterize Protein-Protein Interactions by In-cell NMR Spectroscopy.
Related Articles Using Singular Value Decomposition to Characterize Protein-Protein Interactions by In-cell NMR Spectroscopy.
Chembiochem. 2014 Apr 1;
Authors: Majumder S, Demott CM, Burz DS, Shekhtman A
Abstract
Distinct differences between how model proteins interact in-cell and in vitro suggest that the cytosol might have a profound effect in modulating protein-protein and/or protein-ligand interactions that are not observed in vitro. Analyses of in-cell NMR spectra of target proteins interacting with physiological partners are further complicated by low signal-to-noise ratios, and the long overexpression times used in protein-protein interaction studies may lead to changes in the in-cell spectra over the course of the experiment. To unambiguously resolve the principal binding mode between two interacting species against the dynamic cellular background, we analyzed in-cell spectral data of a target protein over the time course of overexpression of its interacting partner by using single-value decomposition (SVD). SVD differentiates between concentration-dependent and concentration-independent events and identifies the principal binding mode between the two species. The analysis implicates a set of amino acids involved in the specific interaction that differs from previous NMR analyses but is in good agreement with crystallographic data.
PMID: 24692227 [PubMed - as supplied by publisher]
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