Skip to main content
Log in

In vitro preparation of amelogenin nanoparticles carrying nucleic acids

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Amelogenin, a matrix protein involved in biomineralization of enamel, can self-assemble to form nanospheres in a pH-dependent manner. Nucleic acids (single-stranded, double-stranded, and plasmid DNA, as well as RNA) could be co-precipitated with amelogenin, demonstrating a strong binding of nucleic acids to amelogenin. The amounts of co-precipitated nucleic acids were analyzed and binding levels upto 90 μg DNA/mg amelogenin was achieved. The co-precipitation could also be carried out in a bacterial cell homogenate, and no bacterial proteins were found in the amelogenin aggregates, suggesting specificity for nucleic acid binding. Dynamic light scattering showed that amelogenin nanosphere structure is maintained upon DNA binding with an upto 2.6 nm increase in diameter. The reported binding of nucleic acids to amelogenin can be explored practically for nucleic acid separation.

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

Similar content being viewed by others

References

  • Bonde JS, Bulow L (2012) Use of human amelogenin in molecular encapsulation for the design of pH responsive microparticles. BMC Biotechnol 12:25

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Delak K, Harcup C, Lakshminarayanan R, Sun Z, Fan Y, Moradian-Oldak J, Evans JS (2009) The tooth enamel protein, porcine amelogenin, is an intrinsically disordered protein with an extended molecular configuration in the monomeric form. Biochemistry 48:2272–2281

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mandelkern M, Elias JG, Eden D, Crothers DM (1981) The dimensions of DNA in solution. J Mol Biol 152:153–161

    Article  CAS  PubMed  Google Scholar 

  • Margolis HC, Beniash E, Fowler CE (2006) Role of macromolecular assembly of enamel matrix proteins in enamel formation. J Dent Res 85:775–793

    Article  CAS  PubMed  Google Scholar 

  • Moradian-Oldak J, Jimenez I, Maltby D, Fincham AG (2001) Controlled proteolysis of amelogenins reveals exposure of both carboxy- and amino-terminal regions. Biopolymers 58:606–616

    Article  CAS  PubMed  Google Scholar 

  • Moradian-Oldak J, Bouropoulos N, Wang L, Gharakhanian N (2002) Analysis of self-assembly and apatite binding properties of amelogenin proteins lacking the hydrophilic C-terminal. Matrix Biol 21:197–205

    Article  CAS  PubMed  Google Scholar 

  • Orberg ML, Schillen K, Nylander T (2007) Dynamic light scattering and fluorescence study of the interaction between double-stranded DNA and poly(amido amine) dendrimers. Biomacromolecules 8:1557–1563

    Article  PubMed  Google Scholar 

  • Sebastianelli A, Sen T, Bruce IJ (2008) Extraction of DNA from soil using nanoparticles by magnetic bioseparation. Lett Appl Microbiol 46:488–491

    Article  CAS  PubMed  Google Scholar 

  • Svensson Bonde J, Bulow L (2012) One-step purification of recombinant human amelogenin and use of amelogenin as a fusion partner. PLoS One 7:e33269

    Article  PubMed Central  PubMed  Google Scholar 

  • Tan J, Leung W, Moradian-Oldak J, Zeichner-David M, Fincham AG (1998) The pH dependent amelogenin solubility and its biological significance. Connect Tissue Res 38:215–221 discussion 241–216

    Article  CAS  PubMed  Google Scholar 

  • Tian H, Chen J, Chen X (2013) Nanoparticles for gene delivery. Small 9:2034–2044

    Article  CAS  PubMed  Google Scholar 

  • Tsoi M, Do TT, Tang V, Aguilera JA, Perry CC, Milligan JR (2010) Characterization of condensed plasmid DNA models for studying the direct effect of ionizing radiation. Biophys Chem 147:104–110

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Uskokovic V, Castiglione Z, Cubas P, Zhu L, Li W, Habelitz S (2010) Zeta-potential and particle size analysis of human amelogenins. J Dent Res 89:149–153

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Uskokovic V, Odsinada R, Djordjevic S, Habelitz S (2011) Dynamic light scattering and zeta potential of colloidal mixtures of amelogenin and hydroxyapatite in calcium and phosphate rich ionic milieus. Arch Oral Biol 56:521–532

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhang X, Ramirez BE, Liao X, Diekwisch TG (2011) Amelogenin supramolecular assembly in nanospheres defined by a complex helix-coil-PPII helix 3D-structure. PLoS One 6:e24952

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhou Z, Kadam US, Irudayaraj J (2013) One-stop genomic DNA extraction by salicylic acid-coated magnetic nanoparticles. Anal Biochem 442:249–252

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Johan Bonde.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bonde, J., Bülow, L. In vitro preparation of amelogenin nanoparticles carrying nucleic acids. Biotechnol Lett 36, 1349–1357 (2014). https://doi.org/10.1007/s10529-014-1492-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-014-1492-7

Keywords

Navigation