One-step Purification of Recombinant Human Amelogenin and Use of Amelogenin As a Fusion Partner

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2012 Mar 24

PMID 22442680

Citations 17

Authors

Johan Svensson Bonde

Leif Bulow

Affiliations

Soon will be listed here.

Abstract

Amelogenin is an extracellular protein first identified as a matrix component important for formation of dental enamel during tooth development. Lately, amelogenin has also been found to have positive effects on clinical important areas, such as treatment of periodontal defects, wound healing, and bone regeneration. Here we present a simple method for purification of recombinant human amelogenin expressed in Escherichia coli, based on the solubility properties of amelogenin. The method combines cell lysis with recovery/purification of the protein and generates a >95% pure amelogenin in one step using intact harvested cells as starting material. By using amelogenin as a fusion partner we could further demonstrate that the same method also be can explored to purify other target proteins/peptides in an effective manner. For instance, a fusion between the clinically used protein PTH (parathyroid hormone) and amelogenin was successfully expressed and purified, and the amelogenin part could be removed from PTH by using a site-specific protease.

Citing Articles

Recombinant human enamelin produced in Escherichia coli promotes mineralization in vitro.

Halablab M, Wallman L, Bonde J BMC Biotechnol. 2024; 24(1):48.

PMID: 38982413 PMC: 11234762. DOI: 10.1186/s12896-024-00875-0.

Sodium/(calcium + potassium) exchanger NCKX4 optimizes KLK4 activity in the enamel matrix microenvironment to regulate ECM modeling.

Chan B, Cheng I, Rozita J, Gorshteyn I, Huang Y, Shaffer I Front Physiol. 2023; 14:1116091.

PMID: 36814474 PMC: 9939835. DOI: 10.3389/fphys.2023.1116091.

Residue-Specific Insights into the Intermolecular Protein-Protein Interfaces Driving Amelogenin Self-Assembly in Solution.

Buchko G, Mergelsberg S, Tarasevich B, Shaw W Biochemistry. 2022; 61(24):2909-2921.

PMID: 36456190 PMC: 10286517. DOI: 10.1021/acs.biochem.2c00522.

Amyloid-like amelogenin nanoribbons template mineralization via a low-energy interface of ion binding sites.

Akkineni S, Zhu C, Chen J, Song M, Hoff S, Bonde J Proc Natl Acad Sci U S A. 2022; 119(19):e2106965119.

PMID: 35522709 PMC: 9172371. DOI: 10.1073/pnas.2106965119.

WDR72 regulates vesicle trafficking in ameloblasts.

Katsura K, Nakano Y, Zhang Y, Shemirani R, Li W, Den Besten P Sci Rep. 2022; 12(1):2820.

PMID: 35181734 PMC: 8857301. DOI: 10.1038/s41598-022-06751-1.

References

Mirastschijski U, Konrad D, Lundberg E, Lyngstadaas S, Jorgensen L, Agren M . Effects of a topical enamel matrix derivative on skin wound healing. Wound Repair Regen. 2004; 12(1):100-8. DOI: 10.1111/j.1067-1927.2004.012117.x. View

Swanson E, Fong H, Foster B, Paine M, Gibson C, Snead M . Amelogenins regulate expression of genes associated with cementoblasts in vitro. Eur J Oral Sci. 2006; 114 Suppl 1:239-43. DOI: 10.1111/j.1600-0722.2006.00321.x. View

Svensson J, Andersson C, Reseland J, Lyngstadaas P, Bulow L . Histidine tag fusion increases expression levels of active recombinant amelogenin in Escherichia coli. Protein Expr Purif. 2006; 48(1):134-41. DOI: 10.1016/j.pep.2006.01.005. View

Veis A . Amelogenin gene splice products: potential signaling molecules. Cell Mol Life Sci. 2003; 60(1):38-55. PMC: 11138844. DOI: 10.1007/s000180300003. View

Moradian-Oldak J, Du C, Falini G . On the formation of amelogenin microribbons. Eur J Oral Sci. 2006; 114 Suppl 1:289-96. DOI: 10.1111/j.1600-0722.2006.00285.x. View

Lakshminarayanan R, Bromley K, Lei Y, Snead M, Moradian-Oldak J . Perturbed amelogenin secondary structure leads to uncontrolled aggregation in amelogenesis imperfecta mutant proteins. J Biol Chem. 2010; 285(52):40593-603. PMC: 3003358. DOI: 10.1074/jbc.M110.131136. View

Kauvar A, Thoma D, Carnes D, Cochran D . In vivo angiogenic activity of enamel matrix derivative. J Periodontol. 2010; 81(8):1196-201. DOI: 10.1902/jop.2010.090441. View

Fang P, Conway J, Margolis H, Simmer J, Beniash E . Hierarchical self-assembly of amelogenin and the regulation of biomineralization at the nanoscale. Proc Natl Acad Sci U S A. 2011; 108(34):14097-102. PMC: 3161614. DOI: 10.1073/pnas.1106228108. View

Nagano T, Iwata T, Ogata Y, Tanabe T, Gomi K, Fukae M . Effect of heat treatment on bioactivities of enamel matrix derivatives in human periodontal ligament (HPDL) cells. J Periodontal Res. 2004; 39(4):249-56. DOI: 10.1111/j.1600-0765.2004.00733.x. View

10.

Zeichner-David M, Chen L, Hsu Z, Reyna J, Caton J, Bringas P . Amelogenin and ameloblastin show growth-factor like activity in periodontal ligament cells. Eur J Oral Sci. 2006; 114 Suppl 1:244-53. DOI: 10.1111/j.1600-0722.2006.00322.x. View

11.

Fincham A, Moradian-Oldak J, Simmer J . The structural biology of the developing dental enamel matrix. J Struct Biol. 1999; 126(3):270-99. DOI: 10.1006/jsbi.1999.4130. View

12.

Fincham A, Moradian-Oldak J, Simmer J, Sarte P, Lau E, Diekwisch T . Self-assembly of a recombinant amelogenin protein generates supramolecular structures. J Struct Biol. 1994; 112(2):103-9. DOI: 10.1006/jsbi.1994.1011. View

13.

Yuan K, Chen C, Lin M . Enamel matrix derivative exhibits angiogenic effect in vitro and in a murine model. J Clin Periodontol. 2003; 30(8):732-8. DOI: 10.1034/j.1600-051x.2003.00413.x. View

14.

Gestrelius S, Andersson C, Lidstrom D, Hammarstrom L, Somerman M . In vitro studies on periodontal ligament cells and enamel matrix derivative. J Clin Periodontol. 1997; 24(9 Pt 2):685-92. DOI: 10.1111/j.1600-051x.1997.tb00250.x. View

15.

Moradian-Oldak J . Amelogenins: assembly, processing and control of crystal morphology. Matrix Biol. 2001; 20(5-6):293-305. DOI: 10.1016/s0945-053x(01)00154-8. View

16.

Zhu L, Uskokovic V, Le T, DenBesten P, Huang Y, Habelitz S . Altered self-assembly and apatite binding of amelogenin induced by N-terminal proline mutation. Arch Oral Biol. 2010; 56(4):331-6. PMC: 3427771. DOI: 10.1016/j.archoralbio.2010.10.017. View

17.

Viswanathan H, Berry J, Foster B, Gibson C, Li Y, Kulkarni A . Amelogenin: a potential regulator of cementum-associated genes. J Periodontol. 2003; 74(10):1423-31. DOI: 10.1902/jop.2003.74.10.1423. View

18.

Zeichner-David M . Is there more to enamel matrix proteins than biomineralization?. Matrix Biol. 2001; 20(5-6):307-16. DOI: 10.1016/s0945-053x(01)00155-x. View

19.

Heijl L, Heden G, Svardstrom G, Ostgren A . Enamel matrix derivative (EMDOGAIN) in the treatment of intrabony periodontal defects. J Clin Periodontol. 1997; 24(9 Pt 2):705-14. DOI: 10.1111/j.1600-051x.1997.tb00253.x. View

20.

Lyngstadaas S, Wohlfahrt J, Brookes S, Paine M, Snead M, Reseland J . Enamel matrix proteins; old molecules for new applications. Orthod Craniofac Res. 2009; 12(3):243-53. PMC: 2825346. DOI: 10.1111/j.1601-6343.2009.01459.x. View