Ferritin from the haemolymph of adult ants: an extraction method for characterization and a ferromagnetic study

  • Eliane Wajnberg
  • Odivaldo C. Alves
  • Jonas Perales
  • Surza Lucia G. da Rocha
  • André Teixeira Ferreira
  • Luiz Cláudio Cameron
  • Darci M. S. Esquivel
  • Maria de Lourdes Barriviera
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Abstract

Ferritin has been studied in many animals, plants and bacteria. The main functions of ferritin in mammals are iron concentration and stabilization, protection against oxidants and iron storage for later developmental or iron-dependent activities. Although insect ferritin plays a key role in iron transport, only a few studies to date have examined its properties and function. Ferritin isolation from the haemolymph of adult Camponotus sericeiventris ants involved heating at 75 °C, followed by protein fractionation with 3.2 M KBr gradients and ferritin sedimentation with KBr. Protein identification was performed using high-resolution proteomics techniques. SDS-PAGE revealed three subunits with molecular weights (MW) of 26, 28 and 31 kDa. Native PAGE indicated a MW higher than 669 kDa. Proteomic analysis strongly suggested the 26 and 31 kDa bands as F2LCH and F1HCH subunits of ferritin, respectively. Ferromagnetic resonance (FMR) at 100 K showed, at low field, a characteristic broad component of the ferritin iron core, suggesting that its distribution was shifted to values greater than 3000, a higher content than in mammals. The protein yield and MW were comparable to those reported in other studies of insects. To the best of our knowledge, this is the first report on ferritin extracted from adult ants to date. These results are discussed on the basis of the protein structure–function relation of secreted insect and mammal ferritins. This purification method will allow the use of magnetic techniques, which are relevant for understanding the role of ferritin in the biomineralization of magnetic nanoparticles in insects.

Keywords

Ferritin Eusocial insect Ants Ferromagnetic resonance (FMR) Iron transport 
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Notes

Acknowledgements

We are grateful to Dr. Jacques H. C. Delabie, do Lab. de Mirmecologia do CEPEC, for his assistance with the taxonomic classification of Camponotus sericeiventris. M. L. Barriviera was financially supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico. We thank the anonymous reviewers for their constructive comments, which helped us to improve the manuscript and Dr. E. Ellis for linguistic revision.

Supplementary material

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Supplementary material 1 (PDF 127 kb)
249_2018_1293_MOESM2_ESM.pdf (61 kb)
Supplementary material 2 (PDF 61 kb)
249_2018_1293_MOESM3_ESM.pdf (51 kb)
Supplementary material 3 (PDF 50 kb)

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Copyright information

© European Biophysical Societies' Association 2018

Authors and Affiliations

  • Eliane Wajnberg
    • 1
  • Odivaldo C. Alves
    • 2
  • Jonas Perales
    • 4
  • Surza Lucia G. da Rocha
    • 4
  • André Teixeira Ferreira
    • 4
  • Luiz Cláudio Cameron
    • 3
  • Darci M. S. Esquivel
    • 1
  • Maria de Lourdes Barriviera
    • 1
  1. 1.Coordenação de Materiais, Física Aplicada e NanociênciasCentro Brasileiro de Pesquisas FísicasRio de JaneiroBrazil
  2. 2.Departamento de Físico-QuímicaUniversidade Federal FluminenseNiteroi, RJBrazil
  3. 3.Laboratório de Bioquímica de ProteínasUniversidade Federal do Estado do Rio de JaneiroRio de Janeiro, RJBrazil
  4. 4.Laboratório de Toxinologia, Instituto Oswaldo CruzFundação Oswaldo CruzRio de Janeiro, RJBrazil

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