Skip to main content
SpringerLink
Log in

Apolipoprotein B binds ferritin by hemin-mediated binding: evidence of direct binding of apolipoprotein B and ferritin to hemin

  • Published:
BioMetals Aims and scope Submit manuscript

Abstract

Apolipoprotein B (apoB) is known to be a ferritin-binding protein. Here we show that apoB binds to ferritin through hemin-mediated binding. Human apoB bound to bovine spleen, horse spleen, and canine liver ferritins, but did not bind to bovine apoferritin, even after incorporation of iron into it. Incubation of apoferritin with hemin resulted in apoB binding with apoferritin at the same level as with holoferritin. In contrast, hemin inhibited binding of apoB to ferritin. Bovine spleen apoferritin bound biotinylated hemin, and hemin inhibited the binding between the apoferritin and biotinylated hemin, suggesting that ferritin binds hemin directly. ApoB and LDL containing apoB bound biotinylated hemin, and their bindings were also inhibited by hemin, but not protoporphyrin IX. These data demonstrate that binding of apoB to ferritin is mediated through ferritin’s binding to hemin, and also that apoB binds hemin directly.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Andrews SC, Arosio P, Bottke W, Briat JF, von Darl M, Harrison PM, Laulhère JP, Levi S, Lobreaux S, Yewdall SJ (1992) Structure, function, and evolution of ferritins. J Inorg Biochem 47:161–174

    Article  PubMed  CAS  Google Scholar 

  • Addison GM, Beamish MR, Hales CN, Hodgkins M, Jacobs A, Llewellin P (1972) An immunoradiometric assay for ferritin in the serum of normal subjects and patients with iron deficiency and iron overload. J Clin Pathol 25:326–329

    Article  PubMed  CAS  Google Scholar 

  • Avrameas S (1969) Coupling of enzymes to proteins with glutaraldehyde. Use of conjugates for the detection antigens and antibodies. Immunochemistry 6:43–52

    Article  PubMed  CAS  Google Scholar 

  • Camejo G, Halberg C, Manschick-Lundin A, Hurt-Camejo E, Rosengren B, Olsson H, Hansson GI, Forsberg GB, Ylhen B (1998) Hemin binding and oxidation of lipoproteins in serum: mechanisms and effect on the interaction of LDL with human macrophages. J Lipid Res 39:755–766

    PubMed  CAS  Google Scholar 

  • Chan L (1992) Apolipoprotein B, the major protein component of triglyceride-rich and low density lipoproteins. J Biol Chem 36:25621–25624

    Google Scholar 

  • Crichton RR, Soruco JA, Roland F, Michaux MA, Gallois B, Précigoux G, Mahy JP, Mansuy D (1997) Remarkable ability of horse spleen apoferritin to demetallate hemin and to metallate protoporphyrin IX as a function of pH. Biochemistry 36:15049–15054

    Article  PubMed  CAS  Google Scholar 

  • Delanghe JR, Langlois MR (2001) Hemopexin: a review of biological aspects and the role in laboratory medicine. Clin Chim Acta 312:13–23

    Article  PubMed  CAS  Google Scholar 

  • Harvey JW, Beutler E (1982) Binding of heme by glutathione S-transferase: a possible role of the erythrocyte enzyme. Blood 60:1227–1230

    PubMed  CAS  Google Scholar 

  • Hevi S, Chuck SL (2003) Ferritins can regulate the secretion of apolipoproteins B. J Biol Chem 278:31924–31929

    Article  PubMed  CAS  Google Scholar 

  • Ishida M, Dohmae N, Shiro Y, Isogai Y (2003) Synthesis of biotinylated heme and its application to panning heme-binding proteins. Anal Biochem 321:138–141

    Article  PubMed  CAS  Google Scholar 

  • Iwahara S, Satoh H, Song DX, Webb J, Burlingame AL, Nagae Y, Muller-Eberhard U (1995) Purification, characterization, and cloning of a heme-binding protein (23 kDa) in rat liver cytosol. Biochemistry 34:13398–13406

    Article  PubMed  CAS  Google Scholar 

  • Iwasaki K, MacKenzie EL, Hailemariam K, Sakamoto K, Tsuji Y (2006) Hemin-mediated regulation of an antioxidant-responsive element of the human ferritin H gene and role of Ref−1 during erythroid differentiation of K562 cells. Mol Cell Biol 26:2845–2856

    Article  PubMed  CAS  Google Scholar 

  • Jung YD, Chay KO, Song DU, Moon JS, Yang SY, Lee MW, Ahn BW (1997) Hemin inhibits lipid peroxidation induced by ascorbate/FeSO4 and 2,2′-azobis−2-amidino-propane hydrochloride (ABAP). Exp Mol Med 29:171–175

    CAS  Google Scholar 

  • Kadir FHA, Moore GR (1990) Bacterial ferritin contains 24 haem groups. FEBS Lett 271:141–143

    Article  PubMed  CAS  Google Scholar 

  • Kane JP, Hardman DA, Paulus HE (1980) Heterogeneity of apolipoprotein B: isolation of a new species from human chylomicrons. Proc Natl Acad Sci USA 77:2465–2469

    Article  PubMed  CAS  Google Scholar 

  • Kakuta K, Orino K, Yamamoto S, Watanabe K (1997) High levels of ferritin and its iron in fetal bovine serum. Comp Biochem Physiol 118A:165–169

    Article  CAS  Google Scholar 

  • Klausner RD, Harford JB (1989) Cis-trans models for post-transcriptional gene regulation. Science 246:870–872

    Article  PubMed  CAS  Google Scholar 

  • Law SW, Grant SM, Higuchi K, Hospattankar A, Lackner K, Lee N, Brewer HB Jr (1986) Human liver apolipoprotein B−100 cDNA: complete nucleic acid and derived amino acid sequence. Proc Natl Acad Sci USA 83:8142–8146

    Article  PubMed  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  • Massover WH (1994) α2-Macroglobulin: a ferritin-binding protein. Ann NY Acad Sci 737:468–471

    Article  PubMed  CAS  Google Scholar 

  • McCord JM (1996) Effects of positive iron status at a cellular level. Nutr Res 54:85–88

    CAS  Google Scholar 

  • Mertens A, Holvoet P (2001) Oxidized LDL and HDL: antagonists in atherothrombosis. FASEB J 15:2073–2084

    Article  PubMed  CAS  Google Scholar 

  • Orino K, Ishiji T, Yamamoto S, Watanabe K (2004) Characterization of bovine serum ferritin-binding proteins. Comp Biochem Physiol 137A:375–381

    CAS  Google Scholar 

  • Orino K, Uehara M, Okano S, Watanabe K (2006) Purification of characterization of canine serum ferritin-binding proteins. Biometals 19:315–322

    Article  PubMed  CAS  Google Scholar 

  • Orino K, Yamamoto S, Watanabe K (1993) Fibrinogen as a ferritin-binding protein in horse plasma. J Vet Med Sci 55:785–787

    PubMed  CAS  Google Scholar 

  • Pang JHS, Jiang MJ, Chen YL, Wang DL, Chu SH, Chu LY (1996) Increased ferritin gene expression in atherosclerotic lesions. J Clin Invest 97:2204–2212

    Article  PubMed  CAS  Google Scholar 

  • Parthasarathy N, Torti SV, Torti FM (2002) Ferritin binds to light chain of human H-kininogen and inhibits kalliklrein-mediated bradykinin release. Biochem J 365:279–286

    Article  PubMed  CAS  Google Scholar 

  • Rashid KA, Hevi S, Chen Y, Cahérec FL, Chuck SL (2002) A proteomic approach identifies proteins in hepatocytes that bind nascent apolipoprotein B. J Biol Chem 277:22010–22017

    Article  PubMed  CAS  Google Scholar 

  • Rossmann MG, Argos P (1975) A comparison of the heme binding pocket in globins and cytochrome b 5. J Biol Chem 250:7525–7532

    PubMed  CAS  Google Scholar 

  • Santambrogio P, Massover WH (1989) Rabbit serum alpha−2-macroglobulin binds to liver ferritin: association causes a heterogeneity of ferritin molecules. Br J Haematol 71:281–290

    PubMed  CAS  Google Scholar 

  • Segrest JP, Jones MK, Loof HD, Dashti N (2001) Structure of apolipoprotein B−100 in low density lipoproteins. J Lipid Res 42:1346–1367

    PubMed  CAS  Google Scholar 

  • Taketani S, Adachi Y, Kohno H, Ikehara S, Tokunaga R, Ishii T (1998) Molecular characterization of a newly identified heme-binding protein induced during differentiation of murine erythroleukemia cells. J Biol Chem 273:31388–31394

    Article  PubMed  CAS  Google Scholar 

  • Theil EC (1987) Ferritin: structure, gene regulation, and cellular function in animals, plants, and microorganisms. Annu Rev Biochem 56:289–315

    Article  PubMed  CAS  Google Scholar 

  • Torti SV, Torti FM (1998) Human H-kininogen is a ferritin-binding protein. J Biol Chem 273:13630–13635

    Article  PubMed  CAS  Google Scholar 

  • Tsutsui K, Mueller GC (1982) A protein with multiple heme-binding sites from rabbit serum. J Biol Chem 257:3925–3931

    PubMed  CAS  Google Scholar 

  • Worwood M, Cragg SJ, Williams AM, Wagstaff M, Jacobs A (1982) The clearance of 131I-human plasma in man. Blood 60:827–833

    PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This study was partially supported by a Grant for Scientific Research (No. S0603) from School of Veterinary Medicine and Animal Sciences, Kitasato University, Japan.

Author information

Authors and Affiliations

  1. Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Kitasato University, Aomori, 034-8628, Japan

    Takenori Seki, Tomoya Kunichika, Kiyotaka Watanabe & Koichi Orino

Authors
  1. Takenori Seki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tomoya Kunichika
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kiyotaka Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Koichi Orino
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Koichi Orino.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Seki, T., Kunichika, T., Watanabe, K. et al. Apolipoprotein B binds ferritin by hemin-mediated binding: evidence of direct binding of apolipoprotein B and ferritin to hemin. Biometals 21, 61–69 (2008). https://doi.org/10.1007/s10534-007-9093-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10534-007-9093-8

Keywords

Search

Navigation