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Substitution of bovine dentine sialoprotein with chondroitin sulfate glycosaminoglycan chains
Author(s) -
Sugars Rachael V.,
Olsson MarieLouise,
Waddington Rachel,
Wendel Mikael
Publication year - 2006
Publication title -
european journal of oral sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.802
H-Index - 93
eISSN - 1600-0722
pISSN - 0909-8836
DOI - 10.1111/j.1600-0722.2006.00271.x
Subject(s) - chemistry , proteoglycan , gel electrophoresis , blot , chondroitin sulfate , glycosaminoglycan , bone sialoprotein , biochemistry , glycan , sodium dodecyl sulfate , polyacrylamide gel electrophoresis , molecular mass , glycoprotein , extracellular matrix , alkaline phosphatase , enzyme , osteocalcin , gene
Dentine sialoprotein (DSP) represents 5–8% of all non‐collagenous proteins present in the tooth, but, together with dentine phosphoprotein, has been shown to be vital for correct tooth formation. Recently, the existence of a highly glycosylated form of porcine DSP has been reported and it was shown to possess glycosaminoglycan (GAG) chains. The current investigation confirms that this is also the case for bovine DSP and has further characterized these carbohydrates. Dentine sialoprotein was purified from bovine dentine extracts by anion exchange chromatography and identified by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE), western blotting, and mass spectroscopy. An increase in molecular mass was observed, from 120 kDa to greater than 250 kDa, with a corresponding rise in anionic strength. Cellulose acetate electrophoresis and western blotting indicated the presence of chondroitin sulfate GAG chains within these dentine fractions. Further examination using sequential digestion with chondroitinase AC and N ‐glycosidase cleaved the samples first to 95 kDa and then to 80 kDa, respectively, confirming a high level of glycosylation. These results support the classification of bovine DSP as a proteoglycan, and that the carbohydrate substitutions may contribute to the functional properties of DSP.