title: Heme concentration dependence and metalloporphyrin inhibition of the system I and II cytochrome c assembly pathways.
abstract: Studies have indicated that specific heme delivery to apocytochrome c is a critical feature of the cytochrome c biogenesis pathways called system I and II. To determine directly the heme requirements of each system, including whether other metal porphyrins can be incorporated into cytochromes c, we engineered Escherichia coli so that the natural system I (ccmABCDEFGH) was deleted and exogenous porphyrins were the sole source of porphyrins (Delta hemA). The engineered E. coli strains that produced recombinant system I (from E. coli) or system II (from Helicobacter) facilitated studies of the heme concentration dependence of each system. Using this exogenous porphyrin approach, it was shown that in system I the levels of heme used are at least fivefold lower than the levels used in system II, providing an important advantage for system I. Neither system could assemble holocytochromes c with other metal porphyrins, suggesting that the attachment mechanism is specific for Fe protoporphyrin. Surprisingly, Zn and Sn protoporphyrins are potent inhibitors of the pathways, and exogenous heme competes with this inhibition. We propose that the targets are the heme binding proteins in the pathways (CcmC, CcmE, and CcmF for system I and CcsA for system II).
authors.affiliation: Department of Biology, Washington University, St. Louis, MO 63130, USA.
authors.forename: Cynthia L
authors.initials: CL
authors.lastname: Richard-Fogal
authors.forename: Elaine R
authors.initials: ER
authors.lastname: Frawley
authors.forename: Robert E
authors.initials: RE
authors.lastname: Feissner
authors.forename: Robert G
authors.initials: RG
authors.lastname: Kranz
mesh_terms: D003574:Cytochrome c Group / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006418:Heme / Q000378:metabolism* / Q000494:pharmacology; D008665:Metalloporphyrins / Q000494:pharmacology*; D008954:Models, Biological; D009154:Mutation; D011524:Protoporphyrins / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D003574:Cytochrome c Group; D029968:Escherichia coli Proteins; D008665:Metalloporphyrins; D011524:Protoporphyrins; C034186:cytochrome C4; D006418:Heme; C028842:N-methylprotoporphyrin IX