Skip to main content

Advertisement

Log in

Design and characterization of a chimeric ferritin with enhanced iron loading and transverse NMR relaxation rate

  • Original Paper
  • Published:
JBIC Journal of Biological Inorganic Chemistry Aims and scope Submit manuscript

Abstract

This paper describes the design and characterization of a novel ferritin chimera. The iron storage protein ferritin forms a paramagnetic ferrihydrite core. This biomineral, when placed in a magnetic field, can decrease the transverse NMR relaxation times (T 2 and T 2*) of nearby mobile water protons. Ferritin nucleic acid constructs have recently been studied as “probeless” magnetic resonance imaging (MRI) reporters. Following reporter expression, ferritin sequesters endogenous iron and imparts hypointensity to T 2- and T 2*-weighted images in an amount proportional to the ferritin iron load. Wild-type ferritin consists of various ratios of heavy H and light L subunits, and their ratio affects ferritin’s stability and iron storage capacity. We report a novel chimeric ferritin with a fixed subunit stoichiometry obtained by fusion of the L and the H subunits (L*H and H*L) using a flexible linker. We characterize these supramolecular ferritins expressed in human cells, including their iron loading characteristics, hydrodynamic size, subcellular localization, and effect on solvent water T 2 relaxation rate. Interestingly, we found that the L*H chimera exhibits a significantly enhanced iron loading ability and T 2 relaxation compared to wild-type ferritin. We suggest that the L*H chimera may be useful as a sensitive MRI reporter molecule.

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

Access this article

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
Fig. 7

Similar content being viewed by others

Abbreviations

AdV5:

Adenovirus type 5

L*H, H*L:

Chimeric fusion ferritins

DLS:

Dynamic light scatter

H:

Heavy-chain ferritin

IRP2:

Iron-responsive protein 2

L:

Light-chain ferritin

MRI:

Magnetic resonance imaging

PAGE:

Polyacrylamide gel electrophoresis

R 2 :

Transverse relaxation rate

References

  1. Theil EC (1987) Annu Rev Biochem 56:289–315

    Article  CAS  PubMed  Google Scholar 

  2. Theil E, Matzapetakis M, Liu X (2006) J Biol Inorg Chem 11:803–810

    Article  CAS  PubMed  Google Scholar 

  3. Chasteen ND, Harrison PM (1999) J Struct Biol 126:182–194

    Article  CAS  PubMed  Google Scholar 

  4. Vymazal J, Zak O, Bulte JW, Aisen P, Brooks RA (1996) Magn Reson Med 36:61–65

    Article  CAS  PubMed  Google Scholar 

  5. Cohen B, Dafni H, Meir G, Harmelin A, Neeman M (2005) Neoplasia 7:109–117

    Article  CAS  PubMed  Google Scholar 

  6. Genove G, DeMarco U, Xu HY, Goins WF, Ahrens ET (2005) Nat Med 11:450–454

    Article  CAS  PubMed  Google Scholar 

  7. Cohen B, Ziv K, Plaks V, Israely T, Kalchenko V, Harmelin A, Benjamin LE, Neeman M (2007) Nat Med 13:498–503

    Article  CAS  PubMed  Google Scholar 

  8. Deans AE, Wadghiri YZ, Bernas LM, Yu X, Rutt BK, Turnbull DH (2006) Magn Reson Med 56:51–59

    Article  CAS  PubMed  Google Scholar 

  9. Mills PH, Ahrens ET (2007) Magn Reson Med 57:442–447

    Article  PubMed  Google Scholar 

  10. Vymazal J, Brooks RA, Zak O, McRill C, Shen C, Di Chiro G (1992) Magn Reson Med 27:368–374

    Article  CAS  PubMed  Google Scholar 

  11. Gilad AA, Winnard PT Jr, van Zijl PC, Bulte JW (2007) NMR Biomed 20:275–290

    Article  CAS  PubMed  Google Scholar 

  12. Santambrogio P, Levi S, Cozzi A, Rovida E, Albertini A, Arosio P (1993) J Biol Chem 268:12744–12748

    CAS  PubMed  Google Scholar 

  13. Levi S, Yewdall SJ, Harrison PM, Santambrogio P, Cozzi A, Rovida E, Albertini A, Arosio P (1992) Biochem J 288:591–596

    CAS  PubMed  Google Scholar 

  14. Wilkinson Jt, Di X, Schonig K, Buss JL, Kock ND, Cline JM, Saunders TL, Bujard H, Torti SV, Torti FM (2006) Biochem J 395:501–507

    Article  CAS  PubMed  Google Scholar 

  15. Theil E (2007) Biometals 20:513–521

    Article  CAS  PubMed  Google Scholar 

  16. Cozzi A, Corsi B, Levi S, Santambrogio P, Albertini A, Arosio P (2000) J Biol Chem 275:25122–25129

    Article  CAS  PubMed  Google Scholar 

  17. Guo JH, Juan SH, Aust SD (1998) Biochem Biophys Res Commun 242:39–45

    Article  CAS  PubMed  Google Scholar 

  18. Connor JR, Snyder BS, Arosio P, Loeffler DA, LeWitt P (1995) J Neurochem 65:717–724

    Article  CAS  PubMed  Google Scholar 

  19. Kaur D, Rajagopalan S, Chinta S, Kumar J, Di Monte D, Cherny RA, Andersen JK (2007) Brain Res 1140:188–194

    Article  CAS  PubMed  Google Scholar 

  20. Manders EMM, Verbeek FJ, Aten JA (1993) J Microsc 169:375–382

    Google Scholar 

  21. Williams MA, Harrison PM (1968) Biochem J 110:265–280

    CAS  PubMed  Google Scholar 

  22. Donahue KM, Weisskoff RM, Burstein D (1997) J Magn Reson Imaging 7:102–110

    Article  CAS  PubMed  Google Scholar 

  23. Guo B, Phillips JD, Yu Y, Leibold EA (1995) J Biol Chem 270:21645–21651

    Article  CAS  PubMed  Google Scholar 

  24. Rouault TA (2006) Nat Chem Biol 2:406–414

    Article  CAS  PubMed  Google Scholar 

  25. Mosmann T (1983) J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  26. Levi S, Santambrogio P, Cozzi A, Rovida E, Corsi B, Tamborini E, Spada S, Albertini A, Arosio P (1994) J Mol Biol 238:649–654

    Article  CAS  PubMed  Google Scholar 

  27. Voss P, Horakova L, Jakstadt M, Kiekebusch D, Grune T (2006) Free Radic Res 40:673–683

    Article  CAS  PubMed  Google Scholar 

  28. Lee J, Kim SW, Kim YH, Ahn JY (2002) Biochem Biophys Res Commun 298:225–229

    Article  CAS  PubMed  Google Scholar 

  29. Quintana C, Bellefqih S, Laval JY, Guerquin-Kern JL, Wu TD, Avila J, Ferrer I, Arranz R, Patino C (2006) J Struct Biol 153:42–54

    Article  CAS  PubMed  Google Scholar 

  30. Boyd D, Vecoli C, Belcher DM, Jain SK, Drysdale JW (1985) J Biol Chem 260:11755–11761

    CAS  PubMed  Google Scholar 

  31. Surguladze N, Patton S, Cozzi A, Fried MG, Connor JR (2005) Biochem J 388:731–740

    Article  CAS  PubMed  Google Scholar 

  32. Cai CX, Linsenmayer TF (2001) J Cell Sci 114:2327–2334

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We acknowledge valuable technical assistance from Hongyan Xu and Joseph Suhan. We thank Kevin Hitchens for editorial assistance. We acknowledge support from the National Institutes of Health via grants R01-EB005740, P01-HD047675 and P41-EB001977 and a National Science Foundation Graduate Research Fellowship to B.I. (2007053507).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Eric T. Ahrens.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 123 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Iordanova, B., Robison, C.S. & Ahrens, E.T. Design and characterization of a chimeric ferritin with enhanced iron loading and transverse NMR relaxation rate. J Biol Inorg Chem 15, 957–965 (2010). https://doi.org/10.1007/s00775-010-0657-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00775-010-0657-7

Keywords

Navigation