Quantitative analysis of amelogenin solubility.
Amelogenins are a group of extracellular enamel matrix proteins which are believed to be involved in the regulation of the size and habits of forming enamel crystals. The aim of this study was to compare the solubility properties of several amelogenins at various pH (from 4.0 to 9.0) at constant ionic strength (IS), and to examine the influence of buffer composition, IS, and divalent metal ions (including Ca2+, Mg2+, and Zn2+) on amelogenin solubility. The solubility of the recombinant murine amelogenin ("rM179") was minimum near its isoelectric point and increased rapidly below and above, regardless of buffer composition. A similar trend was observed for the native porcine ("25K") amelogenin. Porcine "23K" amelogenin was only sparingly soluble from pH of 4.0 to 9.0, in contrast to the analogous recombinant "rM166", which was more soluble in acidic solutions. The synthetic amelogenin polypeptide "TRAP" was extremely insoluble, while synthetic LRAP was readily soluble. Porcine "20K" amelogenin solubility increased strikingly as the solution pH was lowered from 7.0 to 6.0. Increasing IS decreased the solubility of rM179. While Zn2+ reduced rM179 solubility, Ca2+ and Mg2+ showed no significant effects. We conclude that the solubility of amelogenin was dependent on the primary structure, solution pH, and IS, and the low solubility of amelogenins under physiological conditions may result from their tendency to form quaternary (aggregate) structures in vivo.
Tan
J
J
Center for Craniofacial Molecular Biology, University of Southern California, School of Dentistry, Los Angeles 90033, USA.
Leung
W
W
Moradian-Oldak
J
J
Zeichner-David
M
M
Fincham
A G
AG
D053523:Amelogenin; D000595:Amino Acid Sequence; D000818:Animals; D002021:Buffers; D002118:Calcium / Q000737:chemistry; D003746:Dental Enamel Proteins / Q000737:chemistry*; D003747:Dental Enamel Solubility; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D013552:Swine; D015032:Zinc / Q000737:chemistry
D053523:Amelogenin; C507200:Amelx protein, mouse; D002021:Buffers; D003746:Dental Enamel Proteins; D011994:Recombinant Proteins; C067432:leucine-rich amelogenin peptide; C084495:tyrosine-rich amelogenin polypeptide; D008274:Magnesium; D015032:Zinc; D002118:Calcium