(528b) Mapping the Protein Interaction Between a Clostridial [FeFe] Hydrogenase and Its Redox Partner, Ferredoxin

Electron transport is a fundamental process underlying energy transfer in central metabolism. As electron transport is largely mediated by protein-protein interactions, understanding and manipulating the interactions between electron-carrying proteins can be used to direct electron flow in metabolic pathways engineered to produce reduced compounds including next-generation fuels like hydrogen. [FeFe] hydrogenases from Clostridia catalyze the reaction combining protons and electrons to produce hydrogen and are of significant interest for the development of hydrogen-producing metabolic pathways. The hydrogenase binds to and receives electrons from an electron carrying protein, ferredoxin. However, the interaction between these hydrogenases and ferredoxins is not well understood.
       A protein-protein docking model (ClusPro) was used to generate a structural model of the interaction between the ferredoxin and hydrogenase. This model was then used to predict surface mutations designed to disrupt specific types of interactions that might be occurring at a protein-protein interface. Here, we will discuss the construction of ferredoxin and hydrogenase mutant libraries and our assessment of the effects of these mutations on the interaction between these proteins. This study will provide new, molecular-level insight into the protein interaction that facilitates electron transfer between the Clostridial hydrogenase and its native redox partner. These studies can be further extended to enhance the interaction between these proteins, thereby enabling the construction of more efficient hydrogen production pathways.