title: The Fox1 ferroxidase of Chlamydomonas reinhardtii: a new multicopper oxidase structural paradigm.
abstract: Multicopper oxidases (MCO) contain at least four copper atoms arrayed in three distinct ligand fields supported by two canonical structural features: (1) multiples of the cupredoxin fold and (2) four unique sequence elements that include the ten histidine and one cysteine ligands to the four copper atoms. Ferroxidases are a subfamily of MCO proteins that contain residues supporting a specific reactivity towards ferrous iron; these MCOs play a vital role in iron metabolism in bacteria, algae, fungi, and mammals. In contrast to the fungal ferroxidases, e.g., Fet3p from Saccharomyces cerevisiae, the mammalian ceruloplasmin (Cp) is twice as large (six vs. three cupredoxin domains) and contains three type 1, or "blue," copper sites. Chlamydomonas reinhardtii expresses a putative ferroxidase, Fox1, which has sequence similarity to human Cp (hCp). Eschewing the standard sequence-based modeling paradigm, we have constructed a function-based model of the Fox1 protein which replicates hCp's six copper-site ligand arrays with an overall root mean square deviation of 1.4 A. Analysis of this model has led also to assignment of motifs in Fox1 that are unique to ferroxidases, the strongest evidence to date that the well-characterized fungal high-affinity iron uptake system is essential to iron homeostasis in green algae. The model of Fox1 also establishes a subfamily of MCO proteins with a noncanonical copper-ligand organization. These diverse structures suggest alternative mechanisms for intramolecular electron transfer and require a new trajectory for the evolution of the MCO superfamily.
authors.affiliation: Department of Biochemistry, School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY 14214, USA.
authors.forename: Alaina J
authors.initials: AJ
authors.lastname: Terzulli
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Kosman
mesh_terms: D000465:Algorithms; D000595:Amino Acid Sequence; D000818:Animals; D001400:Azurin / Q000737:chemistry; D001665:Binding Sites; D002570:Ceruloplasmin / Q000737:chemistry* / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000201:enzymology*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: C050107:cupredoxin; D001400:Azurin; D002570:Ceruloplasmin