title: Abcb10 physically interacts with mitoferrin-1 (Slc25a37) to enhance its stability and function in the erythroid mitochondria.
abstract: Mitoferrin-1 (Mfrn1; Slc25a37), a member of the solute carrier family localized in the mitochondrial inner membrane, functions as an essential iron importer for the synthesis of mitochondrial heme and iron-sulfur clusters in erythroblasts. The biochemistry of Mfrn1-mediated iron transport into the mitochondria, however, is poorly understood. Here, we used the strategy of in vivo epitope-tagging affinity purification and mass spectrometry to investigate Mfrn1-mediated mitochondrial iron homeostasis. Abcb10, a mitochondrial inner membrane ATP-binding cassette transporter highly induced during erythroid maturation in hematopoietic tissues, was found as one key protein that physically interacts with Mfrn1 during mouse erythroleukemia (MEL) cell differentiation. Mfrn1 was shown previously to have a longer protein half-life in differentiated MEL cells compared with undifferentiated cells. In this study, Abcb10 was found to enhance the stabilization of Mfrn1 protein in MEL cells and transfected heterologous COS7 cells. In undifferentiated MEL cells, cotransfected Abcb10 specifically interacts with Mfrn1 to enhance its protein stability and promote Mfrn1-dependent mitochondrial iron importation. The structural stabilization of the Mfrn1-Abcb10 complex demonstrates a previously uncharacterized function for Abcb10 in mitochondria. Furthermore, the binding domain of Mfrn1-Abcb10 interaction maps to the N terminus of Mfrn1. These results suggest the tight regulation of mitochondrial iron acquisition and heme synthesis in erythroblasts is mediated by both transcriptional and posttranslational mechanisms, whereby the high level of Mfrn1 is stabilized by oligomeric protein complexes.
authors.affiliation: Department of Medicine, Hematology Division, Brigham and Women's Hospital, and Hematology-Oncology Division, Children's Hospital Boston, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Wen
authors.initials: W
authors.lastname: Chen
authors.forename: Prasad N
authors.initials: PN
authors.lastname: Paradkar
authors.forename: Liangtao
authors.initials: L
authors.lastname: Li
authors.forename: Eric L
authors.initials: EL
authors.lastname: Pierce
authors.forename: Nathaniel B
authors.initials: NB
authors.lastname: Langer
authors.forename: Naoko
authors.initials: N
authors.lastname: Takahashi-Makise
authors.forename: Brigham B
authors.initials: BB
authors.lastname: Hyde
authors.forename: Orian S
authors.initials: OS
authors.lastname: Shirihai
authors.forename: Diane M
authors.initials: DM
authors.lastname: Ward
authors.forename: Jerry
authors.initials: J
authors.lastname: Kaplan
authors.forename: Barry H
authors.initials: BH
authors.lastname: Paw
mesh_terms: D000818:Animals; D001665:Binding Sites; D001692:Biological Transport; D015153:Blotting, Western; D019556:COS Cells; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D047468:Immunoprecipitation; D007501:Iron / Q000378:metabolism; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism / Q000473:pathology; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism*; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D055550:Protein Stability; D014162:Transfection
chemical_list: D007501:Iron; D026901:Membrane Transport Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C498885:Abcc10 protein, mouse; C509051:mitoferrin protein, mouse