Substitution of bovine dentine sialoprotein with chondroitin sulfate glycosaminoglycan chains.
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.
Sugars
Rachael V
RV
Centre for Oral Biology, Karolinska Institutet, Institute of Odontology, Sweden. rachael.sugars@ki.se
Olsson
Marie-Louise
ML
Waddington
Rachel
R
Wendel
Mikael
M
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002809:Chondroitin Sulfates / Q000032:analysis; D002852:Chromatography, Ion Exchange; D003804:Dentin / Q000737:chemistry* / Q000378:metabolism; D004588:Electrophoresis, Cellulose Acetate; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins / Q000737:chemistry; D006031:Glycosylation; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D012795:Sialoglycoproteins / Q000737:chemistry* / Q000145:classification / Q000378:metabolism
D016326:Extracellular Matrix Proteins; D012795:Sialoglycoproteins; C038142:chondroitin sulfate glycosaminoglycan; D002809:Chondroitin Sulfates