title: Overlapping specificities of the mitochondrial cytochrome c and c1 heme lyases.
abstract: Heme attachment to the apoforms of fungal mitochondrial cytochrome c and c1 requires the activity of cytochrome c and c1 heme lyases (CCHL and CC1HL), which are enzymes with distinct substrate specificity. However, the presence of a single heme lyase in higher eukaryotes is suggestive of broader substrate specificity. Here, we demonstrate that yeast CCHL is active toward the non-cognate substrate apocytochrome c1, i.e. CCHL promotes low levels of apocytochrome c1 conversion to its holoform in the absence of CC1HL. Moreover, that the single human heme lyase also displays a broader cytochrome specificity is evident from its ability to substitute for both yeast CCHL and CC1HL. Multicopy and genetic suppressors of the absence of CC1HL were isolated and their analysis revealed that the activity of CCHL toward cytochrome c1 can be enhanced by: 1) reducing the abundance of the cognate substrate apocytochrome c, 2) increasing the accumulation of CCHL, 3) modifying the substrate-enzyme interaction through point mutations in CCHL or cytochrome c1, or 4) overexpressing Cyc2p, a protein known previously only as a mitochondrial biogenesis factor. Based on the functional interaction of Cyc2p with CCHL and the presence of a putative FAD-binding site in the protein, we hypothesize that Cyc2p controls the redox chemistry of the heme lyase reaction.
mesh_terms: D000483:Alleles; D000818:Animals; D001665:Binding Sites; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism; D003575:Cytochromes c1 / Q000737:chemistry / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry; D005656:Fungal Proteins; D005799:Genes, Dominant; D005947:Glucose / Q000737:chemistry; D005990:Glycerol / Q000494:pharmacology; D006418:Heme / Q000737:chemistry; D006801:Humans; D008190:Lyases / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D008954:Models, Biological; D009154:Mutation; D010084:Oxidation-Reduction; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D013045:Species Specificity; D013379:Substrate Specificity; D014466:Ultraviolet Rays
chemical_list: D005656:Fungal Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D005182:Flavin-Adenine Dinucleotide; D006418:Heme; D012313:RNA; D045304:Cytochromes c; D003575:Cytochromes c1; D008190:Lyases; C031581:cytochrome C synthetase; C095893:CYT2 protein, S cerevisiae; D005947:Glucose; D005990:Glycerol