The enamel protein amelogenin binds to the N-acetyl-D-glucosamine-mimicking peptide motif of cytokeratins.
Amelogenins bind to GlcNAc of the dentine-enamel matrix proteins (Ravindranath, R. M. H., Moradian-Oldak, J., Fincham, A. G. (1999) J. Biol. Chem. 274, 2464-2471). The hypothesis that amelogenins may interact with the peptides that mimic GlcNAc is tested. GlcNAc-mimicking peptide (SFGSGFGGGY) but not its variants with single amino acid substitution at serine, tyrosine, or phenylalanine residues inhibited hemagglutination of amelogenins and the terminal tyrosine-rich amelogenin polypeptide (TRAP). The binding affinity of SFGSGFGGGY to amelogenins was confirmed by dosimetric binding of amelogenins or TRAP with [(3)H]peptide, specific binding in varying concentrations of the peptide, Scatchard plot analysis, and competitive inhibition with the unlabeled peptide. The ability of the peptide or GlcNAc to stoichiometrically inhibit TRAP binding of [(14)C]GlcNAc or [(3)H]peptide indicated that both the peptide and GlcNAc compete for a single binding site. Using different fragments of amelogenins, we have identified the peptide-binding motif in amelogenin to be the same as the GlcNAc-binding "amelogenin trityrosyl motif peptide." The GlcNAc-mimicking peptide failed to bind to the amelogenin trityrosyl motif peptide when the tyrosyl residues were substituted with phenylalanine or when the third proline was replaced with threonine, as in some cases of human X-linked amelogenesis imperfecta. This study documents that molecular mimicry may play a role in stability and organization of amelogenin during amelogenesis.
Ravindranath
R M
RM
Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, Los Angeles, California 90033, USA. rravindr@hsc.usc.edu
Tam
W Y
WY
Nguyen
P
P
Fincham
A G
AG
D000117:Acetylglucosamine / Q000737:chemistry*; D053523:Amelogenin; D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D003746:Dental Enamel Proteins / Q000737:chemistry* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D008040:Genetic Linkage; D006388:Hemagglutinins / Q000378:metabolism; D006801:Humans; D007633:Keratins / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry; D010649:Phenylalanine / Q000737:chemistry; D011392:Proline / Q000737:chemistry; D011485:Protein Binding; D012694:Serine / Q000737:chemistry; D013997:Time Factors; D014443:Tyrosine / Q000737:chemistry; D014960:X Chromosome / Q000235:genetics
D053523:Amelogenin; C507200:Amelx protein, mouse; D003746:Dental Enamel Proteins; D006388:Hemagglutinins; D010455:Peptides; C084495:tyrosine-rich amelogenin polypeptide; D014443:Tyrosine; D012694:Serine; D010649:Phenylalanine; D007633:Keratins; D011392:Proline; D000117:Acetylglucosamine