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Proteomic analysis of enamel matrix using a two-dimensional protein fractionation system
- Source :
- European journal of oral sciences. 114
- Publication Year :
- 2006
-
Abstract
- Our objectives in this study were to perform separate proteomic analyses of porcine soft and hard enamel matrices, using the ProteomeLab PF-2D System, to compare the contents of the hard and soft enamel and to identify matrix constituents that are absent from the early maturation stage. Developing first permanent molars were dissected from 6-month-old pigs. Both immature and mature enamel samples were obtained by scraping the secretory-stage (soft) and maturation-stage (hard) enamel, respectively. Enamel matrix samples were sequentially extracted and fractionated with 50 mM phosphate buffer (pH 7.4) and then with 50 mM carbonate buffer (pH 10.8). The neutral enamel extract was separated into four fractions by successive ammonium sulfate precipitations. The alkaline enamel extract was separated into four fractions by ion-exchange chromatography. These eight extracts from both the soft and hard enamel were injected for chromatofocusing. Soft enamel fractions containing constituents absent from the hard enamel were further separated by reverse-phase high-performance liquid chromatography. The major soft enamel constituents absent from the hard enamel were acidic glycoproteins, corresponding to the 32-kDa enamelin, and the 29-, 27-, 15-, 13-, 8- and 6-kDa C-terminal fragments of ameloblastin. Loss of these glycoproteins is associated with a post-transition increase in enamel mineralization.
- Subjects :
- Molar
Ammonium sulfate
Proteome
Swine
Carbonates
Mineralogy
Fractionation
Buffers
Chemical Fractionation
Phosphates
chemistry.chemical_compound
stomatognathic system
Dental Enamel Proteins
Amelogenesis
Animals
Isoelectric Point
Dental Enamel
General Dentistry
Chromatography, High Pressure Liquid
Glycoproteins
Chromatography
Enamel paint
Chromatofocusing
Tooth Germ
Hydrogen-Ion Concentration
Chromatography, Ion Exchange
Peptide Fragments
stomatognathic diseases
Enamel mineralization
Isoelectric point
chemistry
Ammonium Sulfate
visual_art
visual_art.visual_art_medium
Electrophoresis, Polyacrylamide Gel
Subjects
Details
- ISSN :
- 09098836
- Volume :
- 114
- Database :
- OpenAIRE
- Journal :
- European journal of oral sciences
- Accession number :
- edsair.doi.dedup.....438b2b338740eb9a4befb3f0a002cfd2