title: Cytochrome c maturation. The in vitro reactions of horse heart apocytochrome c and Paracoccus dentrificans apocytochrome c550 with heme.
abstract: C-type cytochromes are characterized by having the heme moiety covalently attached via thioether bonds between the heme vinyl groups and the thiols of conserved cysteine residues of the polypeptide chain. Previously, we have shown the in vitro formation of Hydrogenobacter thermophilus cytochrome c(552) (Daltrop, O., Allen, J. W. A., Willis, A. C., and Ferguson, S. J. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 7872-7876). In this work we report that thioether bonds can form spontaneously in vitro between heme and the apocytochromes c from horse heart and Paracoccus denitrificans via b-type cytochrome intermediates. Both apocytochromes, but not the holo forms, bind 8-anilino-1-naphthalenesulfonate, indicating that the apoproteins each have an affinity for a hydrophobic ligand. Furthermore, for both apocytochromes c an intramolecular disulfide can form between the cysteines of the CXXCH motif that is characteristic of c-type cytochromes. In vitro reaction of these apocytochromes c with heme to yield holocytochromes c, and the tendency to form a disulfide, have implications for the different systems responsible for cytochrome c maturation in vivo in various organisms.
authors.affiliation: Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom.
authors.forename: Oliver
authors.initials: O
authors.lastname: Daltrop
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Ferguson
mesh_terms: D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D003545:Cysteine; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism*; D045304:Cytochromes c; D006418:Heme / Q000737:chemistry / Q000378:metabolism*; D006736:Horses; D009206:Myocardium / Q000201:enzymology; D010231:Paracoccus denitrificans / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D001059:Apoproteins; D003574:Cytochrome c Group; C066265:apocytochrome c550, Paracoccus denitrificans; D006418:Heme; D045304:Cytochromes c; D003545:Cysteine