Stability and apoptotic activity of recombinant human cytochrome c

Alina Olteanu, Chetan N. Patel, Matthew M. Dedmon, Scott Kennedy, Michael W. Linhoff, Camille M. Minder, Patrick R. Potts, Mohanish Deshmukh, Gary J. Pielak

Research output: Contribution to journalArticlepeer-review

60 Scopus citations


An efficient system for producing human cytochrome c variants is important to help us understand the roles of this protein in biological processes relevant to human diseases including apoptosis and oxidative stress. Here, we describe an Escherichia coli expression system for producing recombinant human cytochrome c. We also characterize the structure, stability, and function of the protein and show its utility for studying apoptosis. Yields of greater than 8mg of pure protein per liter culture were attained. Circular dichroism spectropolarimetry studies show that the secondary and tertiary structures of the human protein are nearly identical to those of the horse protein, but the human protein is more stable than other eukaryotic cytochromes c. Furthermore, recombinant human cytochrome c is capable of inducing caspase-3 activity in a cell-free caspase activation assay. We use data from this assay along with data from the literature to define the apaf-1 binding site on human cytochrome c.

Original languageEnglish (US)
Pages (from-to)733-740
Number of pages8
JournalBiochemical and Biophysical Research Communications
Issue number3
StatePublished - Dec 19 2003


  • Apoptosis
  • Circular dichroism
  • Cytochrome c
  • Protein stability

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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