title: Amelogenin-enamelin association in phosphate-buffered saline.
abstract: The structures and interactions among macromolecules in the enamel extracellular matrix play vital roles in regulating hydroxyapatite crystal nucleation, growth, and maturation. We used dynamic light scattering (DLS), circular dichroism (CD), fluorescence spectroscopy, and transmission electron microscopy (TEM) to investigate the association of amelogenin and the 32-kDa enamelin, at physiological pH 7.4, in phosphate-buffered saline (PBS). The self-assembly behavior of amelogenin (rP148) was altered following addition of the 32-kDa enamelin. Dynamic light scattering revealed a trend for a decrease in aggregate size in the solution following the addition of enamelin to amelogenin. A blue-shift and intensity increase of the ellipticity minima of rP148 in the CD spectra upon the addition of the 32-kDa enamelin, suggest a direct interaction between the two proteins. In the fluorescence spectra, the maximum emission of rP148 was red-shifted from 335 to 341 nm with a marked intensity increase in the presence of enamelin as a result of complexation of the two proteins. In agreement with DLS data, TEM imaging showed that the 32-kDa enamelin dispersed the amelogenin aggregates into oligomeric particles and stabilized them. Our study provides novel insights into understanding the possible cooperation between enamelin and amelogenin in macromolecular co-assembly and in controlling enamel mineral formation.
authors.affiliation: Center for Craniofacial Molecular Biology, Herman Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90033, USA.
authors.forename: Xiudong
authors.initials: X
authors.lastname: Yang
authors.forename: Daming
authors.initials: D
authors.lastname: Fan
authors.forename: Shibi
authors.initials: S
authors.lastname: Mattew
authors.forename: Janet
authors.initials: J
authors.lastname: Moradian-Oldak
mesh_terms: D000566:Amelogenesis; D053523:Amelogenin / Q000378:metabolism*; D000818:Animals; D002130:Calcium Phosphates / Q000737:chemistry; D002942:Circular Dichroism; D003460:Crystallization; D003746:Dental Enamel Proteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D008970:Molecular Weight; D011485:Protein Binding; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000378:metabolism; D012542:Scattering, Radiation; D012965:Sodium Chloride; D013050:Spectrometry, Fluorescence; D034421:Sus scrofa; D014074:Tooth Calcification
chemical_list: D053523:Amelogenin; D002130:Calcium Phosphates; D003746:Dental Enamel Proteins; D046911:Macromolecular Substances; D011994:Recombinant Proteins; C519480:amorphous calcium phosphate; C038456:tuftelin; C022045:octacalcium phosphate; D012965:Sodium Chloride