title: Characterization of recombinant horse cytochrome c synthesized with the assistance of Escherichia coli cytochrome c maturation factors.
abstract: Cytochromes c are characterized by the presence of a protoporphyrin IX group covalently attached to the polypeptide via one or two thioether bonds to Cys side chains. The heme attachment process, known as cytochrome c maturation, occurs posttranslationally in the periplasm (for bacterial cytochromes c) or in the mitochondrial intermembrane space (for eukaryotic cytochromes c) through a pathway dependent on the organism. It is demonstrated in this work that a mitochondrial cytochrome c expressed in Escherichia coli that undergoes maturation under control of the E. coli cytochrome c maturation factors achieves a native-like structure and stability. The recombinant protein is characterized spectroscopically (by circular dichroism (CD), absorption, and nuclear magnetic resonance (NMR) spectroscopy) and it is verified that the heme and its environment are indistinguishable from authentic horse cytochrome c. Mass spectrometry reveals that the recombinant protein is not acetylated at the N terminus, however, no significant effect on protein structure or stability is detected as a result.
authors.affiliation: Department of Chemistry, College of Arts and Science, University of Rochester, Rochester, NY 14627-0216, USA.
authors.forename: Jason A
authors.initials: JA
authors.lastname: Kellogg
authors.forename: Kara L
authors.initials: KL
authors.lastname: Bren
mesh_terms: D000818:Animals; D002942:Circular Dichroism; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D006736:Horses; D009682:Magnetic Resonance Spectroscopy; D017319:Photosensitizing Agents / Q000378:metabolism; D011487:Protein Conformation; D011524:Protoporphyrins / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D013053:Spectrophotometry
chemical_list: D003574:Cytochrome c Group; D017319:Photosensitizing Agents; D011524:Protoporphyrins; D011994:Recombinant Proteins; C028025:protoporphyrin IX