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

被引:46
作者
Abdullaev, ZK
Bodrova, ME
Chernyak, BV
Dolgikh, DA
Kluck, RM
Perverzev, MO
Arseniev, AS
Efremov, RG
Kirpichnikov, MP
Mokhova, EN
Newmeyer, DD
Roder, H
Skulachev, VP [1 ]
机构
[1] Moscow MV Lomonosov State Univ, AN Belozersky Inst Physicochem Biol, Dept Bioenerget, Moscow 119899, Russia
[2] MM Shemyakin & YA Ovchinnikov Inst Bioorgan Chem, Lab Spectral Anal, Moscow 177871, Russia
[3] La Jolla Inst Allergy & Immunol, Div Cellular Immunol, San Diego, CA 92121 USA
[4] Fox Chase Canc Ctr, Inst Canc Res, Philadelphia, PA 19111 USA
关键词
antioxidant effect; apoptosis; mutagenesis; respiration;
D O I
10.1042/0264-6021:3620749
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 H2O2 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 e-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.
引用
收藏
页码:749 / 754
页数:6
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