Characterization and identification of a human dentin phosphophoryn.
The present study further characterizes an extract from immature, human tooth apicies from which an intact dentin phosphoprotein has been identified. Third molar apicies from developing roots were decalcified in 10% EDTA until Ca2+ was undetectable in the decalcifying solution. The crude extract was run on 7.5% SDS-PAGE and stained with "Stains-All." Four distinct bands were found and the molecular weights were 140, 60, 50, and 34 k. When run on a SDS-PAGE under nonreducing conditions the 60, 50, and 34 k bands were absent. These results suggest that the lower molecular weight bands may be subunits of the larger protein. The extract was then further purified by adding CaCl2 and MgCl2 to precipitate the phosphoprotein. The precipitate was subjected to a DEAE-Sepharose CL6B column and eluted by 0-0.7 M NaCl gradient solution. The amino acid composition of the purified phosphoprotein was determined and the extract was found to be rich in serine and aspartic acid residues. The N-terminal peptide Asp-Asp-Pro was identified. The sequence of the three amino acids is identical to rat incisor phosphoprotein.
Chang
S R
SR
Department of Cariology, Restorative Science, and Endodontics, School of Dentistry, University of Michigan, Ann Arbor, Michigan 48109, U.S.A.
Chiego
D
D
Clarkson
B H
BH
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002122:Calcium Chloride / Q000737:chemistry; D050260:Carbohydrate Metabolism; D003804:Dentin / Q000737:chemistry* / Q000378:metabolism; D004492:Edetic Acid / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007180:Incisor / Q000378:metabolism; D015636:Magnesium Chloride / Q000737:chemistry; D008964:Molar, Third; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D012794:Sialic Acids / Q000378:metabolism; D013194:Staining and Labeling; D014092:Tooth Root / Q000737:chemistry*
D000596:Amino Acids; D010750:Phosphoproteins; D012794:Sialic Acids; C017630:phosphophoryn; D015636:Magnesium Chloride; D004492:Edetic Acid; D019158:N-Acetylneuraminic Acid; D002122:Calcium Chloride