title: Mammalian ferrochelatase, a new addition to the metalloenzyme family.
abstract: A [2Fe-2S] cluster has been detected in mammalian ferrochelatase, the terminal enzyme of the heme biosynthetic pathway. Natural ferrochelatase, purified from mouse livers, and recombinant ferrochelatase, purified from an overproducing strain of Escherichia coli, were investigated by electron paramagnetic resonance (EPR) and Mössbauer spectroscopy. In their reduced forms, both the natural and recombinant ferrochelatases exhibited an identical EPR signal with g values (g = 2.00, 1.93, and 1.90) and relaxation properties typical of [2Fe-2S]+ cluster. Mössbauer spectra of the recombinant ferrochelatase, purified from a strain of E. coli cells transformed with a plasmid encoding murine liver ferrochelatase and grown in 57Fe-enriched medium, demonstrated unambiguously that the cluster is a [2Fe-2S] cluster. No change in the cluster oxidation state was observed during catalysis. The putative protein binding site for the Fe-S cluster in mammalian ferrochelatases is absent from the sequences of the bacterial and yeast enzymes, suggesting a possible role of the [2Fe-2S] center in regulation of mammalian ferrochelatases.
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, University of South Florida, Tampa 33612.
authors.forename: G C
authors.initials: GC
authors.lastname: Ferreira
authors.forename: R
authors.initials: R
authors.lastname: Franco
authors.forename: S G
authors.initials: SG
authors.lastname: Lloyd
authors.forename: A S
authors.initials: AS
authors.lastname: Pereira
authors.forename: I
authors.initials: I
authors.lastname: Moura
authors.forename: J J
authors.initials: JJ
authors.lastname: Moura
authors.forename: B H
authors.initials: BH
authors.lastname: Huynh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli; D005294:Ferrochelatase / Q000737:chemistry* / Q000235:genetics; D007506:Iron-Sulfur Proteins / Q000737:chemistry; D008099:Liver / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D015204:Spectroscopy, Mossbauer
chemical_list: D007506:Iron-Sulfur Proteins; D011994:Recombinant Proteins; D005294:Ferrochelatase