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Home / Equine Research Bank / Biochem J / A cytochrome c mutant with high electron transfer and antiox...
The Biochemical journal2002; 362(Pt 3); 749-754; doi: 10.1042/0264-6021:3620749

A cytochrome c mutant with high electron transfer and antioxidant activities but devoid of apoptogenic effect.

Abstract: A cytochrome c mutant lacking apoptogenic function but competent in electron transfer and antioxidant activities has been constructed. To this end, mutant species of horse and yeast cytochromes c with substitutions in the N-terminal alpha-helix or position 72 were obtained. It was found that yeast cytochrome c was much less effective than the horse protein in activating respiration of rat liver mitoplasts deficient in endogenous cytochrome c as well as in inhibition of H(2)O(2) production by the initial segment of the respiratory chain of intact rat heart mitochondria. The major role in the difference between the horse and yeast proteins was shown to be played by the amino acid residue in position 4 (glutamate in horse, and lysine in yeast; horse protein numbering). A mutant of the yeast cytochrome c containing K4E and some other "horse" modifications in the N-terminal alpha-helix, proved to be (i) much more active in electron transfer and antioxidant activity than the wild-type yeast cytochrome c and (ii), like the yeast cytochrome c, inactive in caspase stimulation, even if added in 400-fold excess compared with the horse protein. Thus this mutant seems to be a good candidate for knock-in studies of the role of cytochrome c-mediated apoptosis, in contrast with the horse K72R, K72G, K72L and K72A mutant cytochromes that at low concentrations were less active in apoptosis than the wild-type, but were quite active when the concentrations were increased by a factor of 2-12.
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Publication Date: 2002-03-07 PubMed ID: 11879204PubMed Central: PMC1222441DOI: 10.1042/0264-6021:3620749Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article is focused on creating a cytochrome c mutant that retains its electron transfer and antioxidant functions, but no longer triggers apoptosis.

Cytochrome c Mutant Research Objective

  • The key objective of this research was to build a cytochrome c mutant that does not have the ability to activate apoptosis (programmed cell death) but still has endothelial transfer and antioxidant functions.

Study Methods and Mutant Creation

  • To achieve the desired objective, researchers experimented with mutant forms of horse and yeast cytochrome c, specifically making modifications to the N-terminal alpha-helix or position 72.
  • The action of yeast cytochrome c, as opposed to horse protein, was found to be less effective in activating respiration in deficient rat liver mitoplasts and in inhibiting H(2)O(2) production in the respiratory chain of rat heart mitochondria.
  • Researchers credited the difference in performance to the amino residue located at position 4. In the horse protein, it is glutamate, whereas in yeast, it is lysine.

Results and Observations

  • A mutant yeast cytochrome c with the K4E mutation in the N-terminal alpha-helix, combined with other “horse” modifications, was found to be much more effective in transferring electrons and antioxidant activity compared to the native yeast cytochrome c.
  • Despite its increased efficiency, this mutant remained as ineffective as the native yeast version in stimulating caspases, even when used in 400 times excess compared to the horse protein.
  • Consequently, this mutant stands as a likely candidate for further studies on cytochrome c-mediated apoptosis.

Comparison with Other Mutants

  • A point of contrast was established with horse mutant cytochromes, K72R, K72G, K72L and K72A that showed inactive to similar apoptotic activity as the wild type at lower concentrations. But their apoptotic activity increased significantly with a 2-12 times rise in concentration.

Cite This Article

APA
Abdullaev ZKh, Bodrova ME, Chernyak BV, Dolgikh DA, Kluck RM, Pereverzev MO, Arseniev AS, Efremov RG, Kirpichnikov MP, Mokhova EN, Newmeyer DD, Roder H, Skulachev VP. (2002). A cytochrome c mutant with high electron transfer and antioxidant activities but devoid of apoptogenic effect. Biochem J, 362(Pt 3), 749-754. https://doi.org/10.1042/0264-6021:3620749

Publication

The Biochemical journal
ISSN: 0264-6021
NlmUniqueID: 2984726R
Country: England
Language: English
Volume: 362
Issue: Pt 3
Pages: 749-754

Researcher Affiliations

Abdullaev, Ziedulla Kh
  • Laboratory of Spectral Analysis, M.M. Shemyakin and Y.A. Ovchinnikov Institute of Bioorganic Chemistry, Moscow 177871, Russia.
Bodrova, Marina E
    Chernyak, Boris V
      Dolgikh, Dmitry A
        Kluck, Ruth M
          Pereverzev, Mikhail O
            Arseniev, Alexander S
              Efremov, Roman G
                Kirpichnikov, Mikhail P
                  Mokhova, Elena N
                    Newmeyer, Donald D
                      Roder, Heinrich
                        Skulachev, Vladimir P

                          MeSH Terms

                          • Amino Acid Sequence
                          • Amino Acid Substitution
                          • Animals
                          • Antioxidants / metabolism
                          • Apoptosis / physiology
                          • Caspases / metabolism
                          • Cytochrome c Group / chemistry
                          • Cytochrome c Group / genetics
                          • Cytochrome c Group / metabolism
                          • Cytochrome c Group / pharmacology
                          • Horses
                          • Kinetics
                          • Mitochondria, Liver / metabolism
                          • Mutagenesis, Site-Directed
                          • Protein Structure, Secondary
                          • Rats
                          • Recombinant Proteins / chemistry
                          • Recombinant Proteins / metabolism
                          • Recombinant Proteins / pharmacology
                          • Saccharomyces cerevisiae / metabolism
                          • Sequence Alignment
                          • Sequence Homology, Amino Acid
                          • Xenopus laevis

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