Comparison of phosphoprotein isolated from mature and immature human tooth roots.
Mature (average patient age = 29.5 yr, closed apical foramen) and immature (average patient age = 17.5 yr, open apical foramen) root shards were placed in dialysis tubing and demineralized to completion using either 10% disodium EDTA plus protease inhibitors or 0.6 N HCl. The demineralized shards were re-extracted (five times) with 0.05 M tris-HCl, 1.0 M NaCl and then collagenase digested. No major differences were observed in chromatograms of extracts, re-extracts or collagenase digests from root shards demineralized in either way. In contrast, chromatograms of immature and mature roots showed qualitative differences. Chromatograms of mature roots demineralized in either way showed broader protein peaks and less organic phosphorus than those from immature tooth roots. A distinct band amid degraded phosphoprotein (150 K) was found in SDS-PAGE gels (7.5%) from EDTA-extracted immature tooth roots but not from mature tooth roots. Electroelution of this band revealed a typical phosphoprotein amino-acid profile containing increased aspartic acid and serine residues. Comparison of the total phosphoprotein and amino acid composition of extracts, re-extracts and collagenase digests revealed that phosphoprotein, serine and to a lesser extent aspartic acid were recovered in greater quantities from immature roots than mature tooth roots. These data suggest that the degree of maturation is crucial to the isolation of an intact phosphoprotein and provides additional evidence that human dentine phosphoprotein undergoes amino acid compositional changes during maturation.
McCurdy
S P
SP
Department of Pediatric Dentistry, University of Connecticut School of Dental Medicine, University of Connecticut Health Center, Farmington 06030.
Clarkson
B H
BH
Feagin
F F
FF
D000293:Adolescent; D000328:Adult; D000596:Amino Acids / Q000032:analysis; D001224:Aspartic Acid / Q000032:analysis; D002852:Chromatography, Ion Exchange; D003804:Dentin / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008964:Molar, Third; D010750:Phosphoproteins / Q000032:analysis / Q000737:chemistry*; D012694:Serine / Q000032:analysis; D014092:Tooth Root / Q000737:chemistry* / Q000254:growth & development*
D000596:Amino Acids; D010750:Phosphoproteins; D001224:Aspartic Acid; D012694:Serine