title: pH triggered self-assembly of native and recombinant amelogenins under physiological pH and temperature in vitro.
abstract: Self-assembly of the extracellular matrix protein amelogenin is believed to play an essential role in regulating the growth and organization of enamel crystals during enamel formation. This study examines the effect of temperature and pH on amelogenin self-assembly under physiological pH conditions in vitro, using dynamic light scattering, turbidity measurements, and transmission electron microscopy. Full-length recombinant amelogenins from mouse (rM179) and pig (rP172) were investigated, along with proteolytic cleavage products (rM166 and native P148) lacking the hydrophilic C-terminus of parent molecules. Results indicated that the self-assembly of full-length amelogenin is primarily triggered by pH in the temperature range from 13 to 37 degrees C and not by temperature. Furthermore, very large assemblies of all proteins studied formed through the rearrangement of similarly sized nanospherical particles, although at different pH values: pH 7.7 (P148), pH 7.5 (rM166), pH 7.2 (rP172), and pH 7.2 (rM179). Structural differences were also observed. The full-length molecules formed apparently tightly connected elongated, high-aspect ratio assemblies comprised of small spheres, while the amelogenin cleavage products appeared as loosely associated spherical particles, suggesting that the hydrophilic C-terminus plays an essential role in higher-order amelogenin assembly. Hence, tightly controlled pH values during secretory amelogenesis may serve to regulate the functions of both full-length and cleaved amelogenins.
authors: Department of Biomineralization, The Forsyth Institute, 140 The Fenway, Boston, MA 02115-3782, USA.
authors: Felicitas B
authors: FB
authors: Wiedemann-Bidlack
authors: Elia
authors: E
authors: Beniash
authors: Yasuo
authors: Y
authors: Yamakoshi
authors: James P
authors: JP
authors: Simmer
authors: Henry C
authors: HC
authors: Margolis
mesh_terms: D053523:Amelogenin / Q000737:chemistry*; D000595:Amino Acid Sequence; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008027:Light; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D017386:Sequence Homology, Amino Acid; D013696:Temperature
chemical_list: D053523:Amelogenin; D011994:Recombinant Proteins
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