title: Effects of phosphorylation on the self-assembly of native full-length porcine amelogenin and its regulation of calcium phosphate formation in vitro.
abstract: The self-assembly of the predominant extracellular enamel matrix protein amelogenin plays an essential role in regulating the growth and organization of enamel mineral during early stages of dental enamel formation. The present study describes the effect of the phosphorylation of a single site on the full-length native porcine amelogenin P173 on self-assembly and on the regulation of spontaneous calcium phosphate formation in vitro. Studies were also conducted using recombinant non-phosphorylated (rP172) porcine amelogenin, along with the most abundant amelogenin cleavage product (P148) and its recombinant form (rP147). Amelogenin self-assembly was assessed using dynamic light scattering (DLS) and transmission electron microscopy (TEM). Using these approaches, we have shown that self-assembly of each amelogenin is very sensitive to pH and appears to be affected by both hydrophilic and hydrophobic interactions. Furthermore, our results suggest that the phosphorylation of the full-length porcine amelogenin P173 has a small but potentially important effect on its higher-order self-assembly into chain-like structures under physiological conditions of pH, temperature, and ionic strength. Although phosphorylation has a subtle effect on the higher-order assembly of full-length amelogenin, native phosphorylated P173 was found to stabilize amorphous calcium phosphate for extended periods of time, in sharp contrast to previous findings using non-phosphorylated rP172. The biological relevance of these findings is discussed.
authors.affiliation: Department of Biomineralization, The Forsyth Institute, Cambridge, MA 02142, USA.
authors.forename: Felicitas B
authors.initials: FB
authors.lastname: Wiedemann-Bidlack
authors.forename: Seo-Young
authors.initials: SY
authors.lastname: Kwak
authors.forename: Elia
authors.initials: E
authors.lastname: Beniash
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Yamakoshi
authors.forename: James P
authors.initials: JP
authors.lastname: Simmer
authors.forename: Henry C
authors.initials: HC
authors.lastname: Margolis
mesh_terms: D053523:Amelogenin / Q000737:chemistry*; D000818:Animals; D002130:Calcium Phosphates / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D046529:Microscopy, Electron, Transmission; D013552:Swine; D013696:Temperature
chemical_list: D053523:Amelogenin; D002130:Calcium Phosphates; C020243:calcium phosphate