Replacement of the invariant lysine 77 by arginine in yeast iso-1-cytochrome c results in enhanced and normal activities in vitro and in vivo.
Oligonucleotide-directed mutagenesis of the yeast Saccharomyces cerevisiae was used to generate an abnormal iso-1-cytochrome c having an Arg-77 replacement of the normal Lys-77; this Lys-77 residue is evolutionarily conserved in most eukaryotic cytochromes c and is trimethylated in fungal and plant cytochromes c. Examination of strains having a single chromosomal copy of the gene encoding the Arg-77 protein indicated that the altered protein was synthesized at the normal rate and that it had normal or near normal activity in vivo. Examination of enzymatic activities in vitro with cytochrome b2, cytochrome c peroxidase, and cytochrome c oxidase indicated that the altered iso-1-cytochrome c has equal or enhanced catalytic efficiencies. Thus, replacement of the evolutionarily conserved residue Lys-77 produces no or only minor effects both in vivo and in vitro.
D000595:Amino Acid Sequence; D001120:Arginine; D002942:Circular Dichroism; D003062:Codon; D003574:Cytochrome c Group / Q000235:genetics / Q000378:metabolism; D003578:Cytochrome-c Peroxidase / Q000378:metabolism; D045304:Cytochromes c; D004271:DNA, Fungal / Q000235:genetics; D003576:Electron Transport Complex IV / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D042844:L-Lactate Dehydrogenase (Cytochrome); D008239:Lysine; D009154:Mutation; D010957:Plasmids; D011487:Protein Conformation; D011489:Protein Denaturation; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D013329:Structure-Activity Relationship; D014170:Transformation, Genetic
C014059:CYC1 protein, S cerevisiae; D003062:Codon; D003574:Cytochrome c Group; D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; D045304:Cytochromes c; D001120:Arginine; D007770:L-Lactate Dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome); D003578:Cytochrome-c Peroxidase; D003576:Electron Transport Complex IV; D008239:Lysine