title: Generation of novel functional metalloproteins via hybrids of cytochrome c and peroxidase.
abstract: The continued interest in protein engineering has led to intense efforts in developing novel stable enzymes, which could not only give boost to industrial and biomedical applications, but also enhance our understanding of the structure-function relationships of proteins. We present here the generation of three hybrid proteins of cytochrome c (cyt c) and peroxidase via structure-based rational mutagenesis of cyt c. Several residues (positions 67, 70, 71 and 80) in the distal heme region of cyt c were mutated to the highly conserved amino acids in the heme pocket of peroxidases. The multiple mutants were found to exhibit high peroxidase activity and conserve the impressive stability of cyt c. We expect that this strategy could be extended to other cases of metalloprotein engineering, and lead to the development of stable and active biocatalysts for industrial uses. Besides, this study also provides insight into the structure-function relationships of hemoproteins.
authors.affiliation: Department of Chemistry & Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Tianlei
authors.initials: T
authors.lastname: Ying
authors.forename: Fangfang
authors.initials: F
authors.lastname: Zhong
authors.forename: Zhong-Hua
authors.initials: ZH
authors.lastname: Wang
authors.forename: Jin
authors.initials: J
authors.lastname: Xie
authors.forename: Xiangshi
authors.initials: X
authors.lastname: Tan
authors.forename: Zhong-Xian
authors.initials: ZX
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D045304:Cytochromes c / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006139:Guaiacol / Q000032:analysis / Q000378:metabolism; D008667:Metalloproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D005656:Fungal Proteins; D008667:Metalloproteins; D011993:Recombinant Fusion Proteins; D006139:Guaiacol; D045304:Cytochromes c; D010544:Peroxidases
keywords: cytochrome c; peroxidase; protein engineering