Comparative analysis of proapoptotic activity of cytochrome c mutants in living cells

horse" hybrid cytochrome c in living WEHI-3 cells. The minimum apoptosis-activating intracellular concentration of horse heart cytochrome c was estimated to be 2.7 +/- 0.5 microM (47 +/- 9 fg/cell). The equieffective concentrations of the K72A-, K72E- and K72L-substituted mutants of cytochrome c were five-, 15- and 70-fold higher. The "yeast --> horse" hybrid created by introducing S2D, K4E, A7K, T8K, and K11V substitutions (horse protein numbering) and deleting five N-terminal residues in yeast cytochrome c did not evoke apoptotic activity in mammalian cells. The apoptotic function of cytochrome c was abolished by the K72W substitution. The K72W-substituted cytochrome c possesses reduced affinity to the apoptotic protease activating factor-1 (Apaf-1) and forms an inactive complex. This mutant is competent as a respiratory-chain electron carrier and well suited for knock-in studies of cytochrome c-mediated apoptosis. -->
ISSN: 1573-675X
1360-8185
Přístupová URL adresa: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c49dbcf238d0c7cc3c036955480c5e38
https://doi.org/10.1007/s10495-005-0366-9
Rights: CLOSED
Přírůstkové číslo: edsair.doi.dedup.....c49dbcf238d0c7cc3c036955480c5e38
Autor: M. V. Astapova, V.I. Dedukhova, Dmitry A. Dolgikh, Alexey V. Feofanov, Kirpichnikov Mp, George V. Sharonov, Vladimir P. Skulachev, Alexander S. Arseniev, Boris V. Chernyak, Olga V. Bocharova
Rok vydání: 2005
Předmět:
Zdroj: Apoptosis. 10:797-808
ISSN: 1573-675X
1360-8185
Popis: A non-traumatic electroporation procedure was developed to load exogenous cytochrome c into the cytoplasm and to study the apoptotic effect of cytochrome c, its K72-substitued mutants and "yeast --> horse" hybrid cytochrome c in living WEHI-3 cells. The minimum apoptosis-activating intracellular concentration of horse heart cytochrome c was estimated to be 2.7 +/- 0.5 microM (47 +/- 9 fg/cell). The equieffective concentrations of the K72A-, K72E- and K72L-substituted mutants of cytochrome c were five-, 15- and 70-fold higher. The "yeast --> horse" hybrid created by introducing S2D, K4E, A7K, T8K, and K11V substitutions (horse protein numbering) and deleting five N-terminal residues in yeast cytochrome c did not evoke apoptotic activity in mammalian cells. The apoptotic function of cytochrome c was abolished by the K72W substitution. The K72W-substituted cytochrome c possesses reduced affinity to the apoptotic protease activating factor-1 (Apaf-1) and forms an inactive complex. This mutant is competent as a respiratory-chain electron carrier and well suited for knock-in studies of cytochrome c-mediated apoptosis.
Databáze: OpenAIRE
Externí odkaz: