title: Amelogenin interacts with cytokeratin-5 in ameloblasts during enamel growth.
abstract: The enamel protein amelogenin binds to GlcNAc (Ravindranath, R. M. H., Moradian-Oldak, R., and Fincham, A.G. (1999) J. Biol. Chem. 274, 2464-2471) and to the GlcNAc-mimicking peptide (GMp) (Ravindranath, R. M. H., Tam, W., Nguyen, P., and Fincham, A. G. (2000) J. Biol. Chem. 275, 39654-39661). The GMp motif in the N-terminal region of the cytokeratin 14 of ameloblasts binds to trityrosyl motif peptide (ATMP) of amelogenin (Ravindranath, R. M. H., Tam, W., Bringas, P., Santos, V., and Fincham, A. G. (2001) J. Biol. Chem. 276, 36586 - 36597). K14 (Type I) pairs with K5 (Type II) in basal epithelial cells; GlcNAc-acylated K5 is identified in ameloblasts. Dosimetric analysis showed the binding affinity of amelogenin to K5 and to GlcNAc-acylated-positive control, ovalbumin. The specific binding of [3H]ATMP with K5 or ovalbumin was confirmed by Scatchard analysis. [3H]ATMP failed to bind to K5 after removal of GlcNAc. Blocking K5 with ATMP abrogates the K5-amelogenin interaction. K5 failed to bind to ATMP when the third proline was substituted with threonine, as in some cases of human X-linked amelogenesis imperfecta or when tyrosyl residues were substituted with phenylalanine. Confocal laser scan microscopic observations on ameloblasts during postnatal (PN) growth of the teeth showed that the K5-amelogenin complex migrated from the cytoplasm to the periphery (on PN day 1) and accumulated at the apical region on day 3. Secretion of amelogenin commences from day 1. K5, similar to K14, may play a role of chaperone during secretion of amelogenin. Upon secretion of amelogenin, K5 pairs with K14. Pairing of K5 and K14 commences on day 3 and ends on day 9. The pairing of K5 and K14 marks the end of secretion of amelogenin.
authors.affiliation: Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, Los Angeles 90033-1004, USA. rravindr@usc.edu
authors.forename: Rajeswari M H
authors.initials: RM
authors.lastname: Ravindranath
authors.forename: Rajam M
authors.initials: RM
authors.lastname: Basilrose
authors.forename: Naren H
authors.initials: NH
authors.lastname: Ravindranath
authors.forename: Bhavapriya
authors.initials: B
authors.lastname: Vaitheesvaran
mesh_terms: D053523:Amelogenin; D000818:Animals; D015153:Blotting, Western; D003743:Dental Enamel / Q000166:cytology*; D003746:Dental Enamel Proteins / Q000378:metabolism*; D006801:Humans; D053569:Keratin-15; D053555:Keratin-5; D007633:Keratins / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D011485:Protein Binding
chemical_list: D053523:Amelogenin; C507200:Amelx protein, mouse; D003746:Dental Enamel Proteins; C503940:KRT5 protein, human; D053569:Keratin-15; D053555:Keratin-5; C492053:Krt15 protein, mouse; D007633:Keratins