title: Design of Heteronuclear Metalloenzymes.
abstract: Heteronuclear metalloenzymes catalyze some of the most fundamentally interesting and practically useful reactions in nature. However, the presence of two or more metal ions in close proximity in these enzymes makes them more difficult to prepare and study than homonuclear metalloenzymes. To meet these challenges, heteronuclear metal centers have been designed into small and stable proteins with rigid scaffolds to understand how these heteronuclear centers are constructed and the mechanism of their function. This chapter describes methods for designing heterobinuclear metal centers in a protein scaffold by giving specific examples of a few heme-nonheme bimetallic centers engineered in myoglobin and cytochrome c peroxidase. We provide step-by-step procedures on how to choose the protein scaffold, design a heterobinuclear metal center in the protein scaffold computationally, incorporate metal ions into the protein, and characterize the resulting metalloproteins, both structurally and functionally. Finally, we discuss how an initial design can be further improved by rationally tuning its secondary coordination sphere, electron/proton transfer rates, and the substrate affinity.
authors: University of Illinois at Urbana-Champaign, Urbana, IL, United States.
authors: A
authors: A
authors: Bhagi-Damodaran
authors: University of Illinois at Urbana-Champaign, Urbana, IL, United States.
authors: P
authors: P
authors: Hosseinzadeh
authors: University of Illinois at Urbana-Champaign, Urbana, IL, United States.
authors: E
authors: E
authors: Mirts
authors: University of Illinois at Urbana-Champaign, Urbana, IL, United States.
authors: J
authors: J
authors: Reed
authors: University of Illinois at Urbana-Champaign, Urbana, IL, United States.
authors: I D
authors: ID
authors: Petrik
authors: University of Illinois at Urbana-Champaign, Urbana, IL, United States. Electronic address: yi-lu@illinois.edu.
authors: Y
authors: Y
authors: Lu
mesh_terms: D002384:Catalysis; D003578:Cytochrome-c Peroxidase / Q000737:chemistry; D006418:Heme / Q000138:chemical synthesis / Q000737:chemistry*; D007477:Ions / Q000737:chemistry; D008667:Metalloproteins / Q000138:chemical synthesis / Q000737:chemistry*; D008670:Metals / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry*; D010084:Oxidation-Reduction; D015202:Protein Engineering / Q000379:methods*
chemical_list: D007477:Ions; D008667:Metalloproteins; D008670:Metals; D009211:Myoglobin; D006418:Heme; D003578:Cytochrome-c Peroxidase
keywords: Biomimetic models; Biosynthetic models; Heme–copper oxidase; Manganese peroxidase; Metalloprotein design; Nitric oxide reductase; Oxygen activation; Protein design; Protein engineering; Secondary sphere interactions