title: Expression and characterization of recombinant human cytochrome c in E. coli.
abstract: Cytochrome c is a heme protein involved in electron transfer, cell apoptosis, and diseases associated with oxidative stress. Here we expressed human cytochrome c in E. coli and purified it to homogeneity with a yield of 10-15 mg/L. The redox potential of recombinant human cytochrome c was 0.246 V which was measured by cyclic voltammetry. This is similar to that of horse cytochrome c with a value of 0.249 V. The sequential assignment and structural analysis of recombinant human ferrocytochrome c were obtained using multidimensional NMR spectroscopy. On the basis of our NMR studies, the recombinant human cytochrome c produced in E. coli exhibits the same tertiary fold as horse cytochrome c. These results provide evidence that human cytochrome c expressed in E. coli possesses a similar function and structure to that of the horse protein. It is known that cytochrome c plays a role in many human diseases. This study serves as the basis for gaining insight into human diseases by exploring structure and function relationships of cytochrome c to its interacting proteins.
authors.affiliation: Department of Biochemistry, National Cheng Kung University College of Medicine, Tainan 701, Taiwan.
authors.forename: Wen-Yih
authors.initials: WY
authors.lastname: Jeng
authors.forename: Chiu-Yueh
authors.initials: CY
authors.lastname: Chen
authors.forename: Hsien-Chang
authors.initials: HC
authors.lastname: Chang
authors.forename: Woei-Jer
authors.initials: WJ
authors.lastname: Chuang
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003574:Cytochrome c Group / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics; D004563:Electrochemistry; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013053:Spectrophotometry
chemical_list: D003574:Cytochrome c Group; D004274:DNA, Recombinant; D011994:Recombinant Proteins