NMR techniques for characterization of proteins under physiological conditions and peptide-food additive interactions
In the frame of the present proposal atomic level characterization of intrinsically disordered proteins (IDPs) and small molecules will be performed.
In general IDPs are studied at lower temperatures and acidic media; determination of the chemical environments is based on HN-detected 3D measurements. Still, some biologically relevant interactions take place only under physiological conditions. In this case 15N-HSQC spectra may present significant peak broadening due to the proton exchange between the solvent water and the dissociable -NH. Therefore different routes have to be chosen, such as the Hα – detection. In this approach the water signal and the multiple splitting due to isotope labeling are the bottlenecks. In our present study we intend to try experimental techniques related Hα-detection. For this purpose we use the 105-residues long EZH2 IDP.
The other focus will be targeted towards emerging NMR techniques for small molecule studies (fast 2D homo- and heteronuclear measurements, translational diffusion measurements) in the characterization of the interaction between a relevant antimicrobial peptide and a food colorant. So far little is known about these types of AMP interactions, thus understanding their action mechanism will contribute to a better description of their activity.
Andrea Bodor, Ágnes Tantos, Tamás Beke-Somfai