title: Examination of ferrochelatase mutations that cause erythropoietic protoporphyria.
abstract: Ferrochelatase (E.C. 4.99.1.1), the enzyme that catalyzes the terminal step in the heme biosynthetic pathway, is the site of defect in the human inherited disease erythropoietic protoporphyria (EPP). Previously it has been demonstrated that patients with EPP may have missense mutations leading to amino acid substitutions, early chain termination, or exon deletions. While it has been clearly demonstrated that two missense mutations result in lowered enzyme activity, it has never been shown what effect specific exon deletions may have. In the current work, recombinant human ferrochelatase has been engineered to have individual exon deletions corresponding to exons 3 through 11. When expressed in Escherichia coli, none of these possesses significant enzyme activity and all lack the [2Fe-2S] cluster. One of the human missense mutations, F417S, and a series of amino acid replacements at this site (ie, F417W, F417Y, and F417L) were examined. With the exception of F417L, all lacked enzyme activity and did not contain the [2Fe-2S] cluster in vivo or as isolated in vitro.
authors.affiliation: Department of Microbiology, University of Georgia, Athens, GA, USA.
authors.forename: V M
authors.initials: VM
authors.lastname: Sellers
authors.forename: T A
authors.initials: TA
authors.lastname: Dailey
authors.forename: H A
authors.initials: HA
authors.lastname: Dailey
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004926:Escherichia coli; D005091:Exons / Q000235:genetics; D005294:Ferrochelatase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D017354:Point Mutation; D017121:Porphyria, Hepatoerythropoietic / Q000235:genetics*; D046351:Protoporphyria, Erythropoietic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion
chemical_list: D011993:Recombinant Fusion Proteins; D007501:Iron; D005294:Ferrochelatase