title: CcmI subunit of CcmFHI heme ligation complex functions as an apocytochrome c chaperone during c-type cytochrome maturation.
abstract: Cytochrome c maturation (Ccm) is a sophisticated post-translational process. It occurs after translocation of apocytochromes c to the p side of energy transducing membranes and forms stereo-specific thioether bonds between the vinyl groups of heme b (protoporphyrin IX-Fe) and the thiol groups of cysteines at their conserved heme binding sites. In many organisms this process involves up to 10 (CcmABCDEFGHI and CcdA) membrane proteins. One of these proteins is CcmI, which has an N-terminal membrane-embedded domain with two transmembrane helices and a large C-terminal periplasmic domain with protein-protein interaction motifs. Together with CcmF and CcmH, CcmI forms a multisubunit heme ligation complex. How the CcmFHI complex recognizes its apocytochrome c substrates remained unknown. In this study, using Rhodobacter capsulatus apocytochrome c(2) as a Ccm substrate, we demonstrate for the first time that CcmI binds apocytochrome c(2) but not holocytochrome c(2). Mainly the C-terminal portions of both CcmI and apocytochrome c(2) mediate this binding. Other physical interactions via the conserved structural elements in apocytochrome c(2), like the heme ligating cysteines or heme iron axial ligands, are less crucial. Furthermore, we show that the N-terminal domain of CcmI can also weakly bind apocytochrome c(2), but this interaction requires a free thiol group at apocytochrome c(2) heme binding site. We conclude that the CcmI subunit of the CcmFHI complex functions as an apocytochrome c chaperone during the Ccm process used by proteobacteria, archaea, mitochondria of plants and red algae.
authors.affiliation: Department of Biology, University of Pennsylvania Philadelphia, Pennsylvania 19014-6019, USA.
authors.forename: Andreia F
authors.initials: AF
authors.lastname: Verissimo
authors.forename: Honghui
authors.initials: H
authors.lastname: Yang
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Wu
authors.forename: Carsten
authors.initials: C
authors.lastname: Sanders
authors.forename: Fevzi
authors.initials: F
authors.lastname: Daldal
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism*; D045362:Cytochromes c2 / Q000378:metabolism; D000939:Epitopes / Q000378:metabolism; D006418:Heme / Q000378:metabolism*; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D019897:Periplasm / Q000201:enzymology; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D016354:Rhodobacter capsulatus / Q000166:cytology / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D000939:Epitopes; D018832:Molecular Chaperones; D010446:Peptide Fragments; D021122:Protein Subunits; D006418:Heme; D045304:Cytochromes c; D045362:Cytochromes c2