title: Equilibrium Studies of Designed Metalloproteins.
abstract: Complete thermodynamic descriptions of the interactions of cofactors with proteins via equilibrium studies are challenging, but are essential to the evaluation of designed metalloproteins. While decades of studies on protein-protein interaction thermodynamics provide a strong underpinning to the successful computational design of novel protein folds and de novo proteins with enzymatic activity, the corresponding paucity of data on metal-protein interaction thermodynamics limits the success of computational metalloprotein design efforts. By evaluating the thermodynamics of metal-protein interactions via equilibrium binding studies, protein unfolding free energy determinations, proton competition equilibria, and electrochemistry, a more robust basis for the computational design of metalloproteins may be provided. Our laboratory has shown that such studies provide detailed insight into the assembly and stability of designed metalloproteins, allow for parsing apart the free energy contributions of metal-ligand interactions from those of porphyrin-protein interactions in hemeproteins, and even reveal their mechanisms of proton-coupled electron transfer. Here, we highlight studies that reveal the complex interplay between the various equilibria that underlie metalloprotein assembly and stability and the utility of making these detailed measurements.
authors: Brooklyn College, Brooklyn, NY, United States; Ph.D. Programs in Chemistry and Biochemistry, The Graduate Center of the City University of New York, New York, NY, United States. Electronic address: BGibney@brooklyn.cuny.edu.
authors: B R
authors: BR
authors: Gibney
mesh_terms: D001665:Binding Sites; D019295:Computational Biology / Q000379:methods; D004734:Energy Metabolism; D006420:Hemeproteins / Q000737:chemistry*; D008024:Ligands; D008667:Metalloproteins / Q000138:chemical synthesis / Q000737:chemistry*; D008670:Metals / Q000737:chemistry*; D015202:Protein Engineering / Q000379:methods*; D058767:Protein Unfolding; D011522:Protons; D013816:Thermodynamics
chemical_list: D006420:Hemeproteins; D008024:Ligands; D008667:Metalloproteins; D008670:Metals; D011522:Protons
keywords: Electrochemistry; Hemeprotein; Protein design; Thermodynamics