title: Interactions of amelogenin with phospholipids.
abstract: Amelogenin protein has the potential to interact with other enamel matrix proteins, mineral, and cell surfaces. We investigated the interactions of recombinant amelogenin rP172 with small unilamellar vesicles as model membranes, toward the goal of understanding the mechanisms of amelogenin-cell interactions during amelogenesis. Dynamic light scattering (DLS), fluorescence spectroscopy, circular dichroism (CD), and nuclear magnetic resonance (NMR) were used. In the presence of phospholipid vesicles, a blue shift in the Trp fluorescence emission maxima of rP172 was observed (∼334 nm) and the Trp residues of rP172 were inaccessible to the aqueous quencher acrylamide. DLS studies indicated complexation of rP172 and phospholipids, although the possibility of fusion of phospholipids following amelogenin addition cannot be ruled out. NMR and CD studies revealed a disorder-order transition of rP172 in a model membrane environment. Strong fluorescence resonance energy transfer from Trp in rP172 to DNS-bound-phospholipid was observed, and fluorescence polarization studies indicated that rP172 interacted with the hydrophobic core region of model membranes. Our data suggest that amelogenin has ability to interact with phospholipids and that such interactions may play key roles in enamel biomineralization as well as reported amelogenin signaling activities.
authors.affiliation: Center for Craniofacial Molecular Biology, Division of Biomedical Sciences, University of Southern California, Herman Ostrow School of Dentistry of USC, Los Angeles, CA, 90033.
authors.forename: Sowmya Bekshe
authors.initials: SB
authors.lastname: Lokappa
authors.forename: Karthik Balakrishna
authors.initials: KB
authors.lastname: Chandrababu
authors.forename: Kaushik
authors.initials: K
authors.lastname: Dutta
authors.forename: Iva
authors.initials: I
authors.lastname: Perovic
authors.forename: John Spencer
authors.initials: JS
authors.lastname: Evans
authors.forename: Janet
authors.initials: J
authors.lastname: Moradian-Oldak
mesh_terms: D053523:Amelogenin / Q000737:chemistry* / Q000378:metabolism*; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence
chemical_list: D053523:Amelogenin; D010743:Phospholipids
keywords: amelogenin; cell membrane; enamel; intrinsically disordered protein; lipid vesicles