The antimycin binding sites were found to be independent of cytochrome(s) b synthesis in pseudo-wild-type revertants of cytochrome b mutants. These revertants, whose primary mutation is located in the introns of the cob-box gene of mitochondrial DNA, have modified contents of cytochromes b-562 and b-565 and fully functional respiratory chain. 2. Missense mutations in three genetic loci allocated to the exons of the cytochrome b gene, abolish the strong affinity binding of antimycin. 3. It is proposed that the antimycin-binding component is not cytochrome b itself, but interacts with it in such a way that an alteration of the cytochrome b structure affects the antimycin-binding site.