title: Characterization of a single b-type heme, FAD, and metal binding sites in the transmembrane domain of six-transmembrane epithelial antigen of the prostate (STEAP) family proteins.
abstract: Six-transmembrane epithelial antigen of the prostate 3 (Steap3) is the major ferric reductase in developing erythrocytes. Steap family proteins are defined by a shared transmembrane domain that in Steap3 has been shown to function as a transmembrane electron shuttle, moving cytoplasmic electrons derived from NADPH across the lipid bilayer to the extracellular face where they are used to reduce Fe(3+) to Fe(2+) and potentially Cu(2+) to Cu(1+). Although the cytoplasmic N-terminal oxidoreductase domain of Steap3 and Steap4 are relatively well characterized, little work has been done to characterize the transmembrane domain of any member of the Steap family. Here we identify high affinity FAD and iron biding sites and characterize a single b-type heme binding site in the Steap3 transmembrane domain. Furthermore, we show that Steap3 is functional as a homodimer and that it utilizes an intrasubunit electron transfer pathway through the single heme moiety rather than an intersubunit electron pathway through a potential domain-swapped dimer. Importantly, the sequence motifs in the transmembrane domain that are associated with the FAD and metal binding sites are not only present in Steap2 and Steap4 but also in Steap1, which lacks the N-terminal oxidoreductase domain. This strongly suggests that Steap1 harbors latent oxidoreductase activity.
authors.affiliation: From the Departments of Chemistry and Biochemistry and.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Kleven
authors.affiliation: Microbiology and Immunology, Montana State University, Bozeman, Montana 59717.
authors.forename: Mensur
authors.initials: M
authors.lastname: Dlakić
authors.affiliation: From the Departments of Chemistry and Biochemistry and lawrence@chemistry.montana.edu.
authors.forename: C Martin
authors.initials: CM
authors.lastname: Lawrence
mesh_terms: D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism*; D003300:Copper / Q000378:metabolism*; D005182:Flavin-Adenine Dinucleotide / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006418:Heme / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007501:Iron / Q000378:metabolism*; D009249:NADP / Q000235:genetics / Q000378:metabolism*; D055503:Protein Multimerization / Q000502:physiology; D017434:Protein Structure, Tertiary
chemical_list: D000951:Antigens, Neoplasm; D005182:Flavin-Adenine Dinucleotide; D006418:Heme; D009249:NADP; D003300:Copper; D007501:Iron
keywords: Steap1; Steap3; b-type cytochrome; copper; copper transport; cupric reductase; ferric reductase; flavin adenine dinucleotide (FAD); iron; iron metabolism