In an aqueous solution of metalloproteins and ions, the concentration and type of ions have a huge effect on the protein aggregation properties. In this two-year project, a postdoc will use a combination of advanced molecular simulation techniques and an experimental approach involving sophisticated separation techniques to obtain crucial insights into the binding of various ions to metalloproteins. This will provide a deeper understanding of how these type of enzymes function in complex fluids such as dairy products.

Combining computational and analytical chemistry

Molecular dynamics (MD) simulations are particularly well suited for studying ions interacting with proteins and can generate insights at high spatial and temporal resolution. Smart sampling algorithms allow the investigation of rare transitions in molecular systems, closing the gap to experimental time and length scales even further.

This project aims to predict ion binding sites, the affinity of metalloproteins for various ions, and the mechanisms of ion binding to the enzyme, aiding the understanding of the effects of ions on the dimerization and tetramerization properties. These predictions will be tested by asymmetrical-flow field-flow fractionation with multi-angle light scattering detection (AF4-MALS) experiments. This technique is ideal to study aggregation behaviour and to characterize very large and highly complicated self-assemblies of such enzymes in solution.

The experimental data together with the predictions on ion binding sites and the affinity of the enzyme for different ions will help shed light onto the effect of various ions on dimerization and tetramerization properties, conformational changes and on how these processes occur.