Glycosaminoglycans and glycoproteins isolated from rabbit femur.
1. Complex carbohydrate fractions were extracted successively with 40% aqueous EDTA (pH 7.4) and 6M urea (PH 7.8) FROM ACETONE-DRIED bone powder of rabbit femur. 2. The carbohydrate fraction extracted with EDTA (E=Fr) was separated into five fractions,D1approximatelyD5 by DEAE-Dephadex A-50 column chromatography. Chemical and infrared spectral analyses, and enzymatic digestion indicate that D2 contained lessacidic glycoprotein, D3 contained sialoglycoprotein, D4 contained a low sulfated proteokeratan sulfate-like substance, and d5 contained glycoprotein-bound chondroitin sulfate A plus protein-free chondroitin sulfate A. 3. Two fractions, HU-D1 and HU-D2, were isolated from the carbohydrate fraction extracted with urea (HU-Fr) by successive digestion with collagenase [EC 3.4.99.5] and pronase, followed by gel-filtration on Sephadex G-100 and then DEAE-Sephadex A-50 column chromatography. HU-D1 and HU-D2 contained a low sulfated keratan sulfate-like substance linked to peptide and glycopeptide-bound chondroitin sulfated keratan sulfate-like substance linked to peptide and glycopeptide-bound chondroitin sulfate A, respectively. 4. The present findings indicate that rabbit femur contains low sulfated proteokeratan sulfate-like substances with varying sulfate contents and glycoprotein-bound chondroitin sulfate A as the principal glycosaminoglycans. The macromolecules bound more tightly to the tissue contain much more sulfate than the corresponding loosely bound ones.
D000818:Animals; D002807:Chondroitin / Q000032:analysis; D004492:Edetic Acid / Q000494:pharmacology; D005269:Femur / Q000032:analysis*; D006023:Glycoproteins / Q000302:isolation & purification*; D006025:Glycosaminoglycans / Q000302:isolation & purification*; D006601:Hexoses / Q000032:analysis; D008722:Methods; D003012:Microbial Collagenase / Q000494:pharmacology; D011402:Pronase / Q000494:pharmacology; D011817:Rabbits; D013055:Spectrophotometry, Infrared; D013431:Sulfates / Q000032:analysis; D014508:Urea / Q000494:pharmacology
D006023:Glycoproteins; D006025:Glycosaminoglycans; D006601:Hexoses; D013431:Sulfates; D014508:Urea; D002807:Chondroitin; D004492:Edetic Acid; D011402:Pronase; D003012:Microbial Collagenase