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J Biol Chem. 2013 Aug 2;288(31):22333-45. doi: 10.1074/jbc.M113.456012. Epub 2013 Jun 19.

Intrinsically disordered enamel matrix protein ameloblastin forms ribbon-like supramolecular structures via an N-terminal segment encoded by exon 5.

Author information

1
Institute of Microbiology, Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic.

Abstract

Tooth enamel, the hardest tissue in the body, is formed by the evolutionarily highly conserved biomineralization process that is controlled by extracellular matrix proteins. The intrinsically disordered matrix protein ameloblastin (AMBN) is the most abundant nonamelogenin protein of the developing enamel and a key element for correct enamel formation. AMBN was suggested to be a cell adhesion molecule that regulates proliferation and differentiation of ameloblasts. Nevertheless, detailed structural and functional studies on AMBN have been substantially limited by the paucity of the purified nondegraded protein. With this study, we have developed a procedure for production of a highly purified form of recombinant human AMBN in quantities that allowed its structural characterization. Using size exclusion chromatography, analytical ultracentrifugation, transmission electron, and atomic force microscopy techniques, we show that AMBN self-associates into ribbon-like supramolecular structures with average widths and thicknesses of 18 and 0.34 nm, respectively. The AMBN ribbons exhibited lengths ranging from tens to hundreds of nm. Deletion analysis and NMR spectroscopy revealed that an N-terminal segment encoded by exon 5 comprises two short independently structured regions and plays a key role in self-assembly of AMBN.

KEYWORDS:

Ameloblastin; Amelogenin; Extracellular Matrix Proteins; Intrinsically Disordered Proteins; Protein Purification; Protein Self-assembly; Tooth

PMID:
23782691
PMCID:
PMC3829324
DOI:
10.1074/jbc.M113.456012
[Indexed for MEDLINE]
Free PMC Article

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