Analysis of the proteoglycans synthesized by corneal explants from embryonic chicken. II. Structural characterization of the keratan sulfate and dermatan sulfate proteoglycans from corneal stroma.
Radioisotopically labeled proteoglycans were isolated from a 4 M guanidine HCl, 2% Triton X-100 extract of corneal stroma from day 18 chicken embryos by anion-exchange chromatography. Two predominant proteoglycans in the sample were separated by octyl-Sepharose chromatography using a gradient elution of detergent in 4 M guanidine HCl. One proteoglycan had an overall mass of approximately 125 kDa, a single dermatan sulfate chain (approximately 85-90% chondroitin 4-sulfate, low iduronate content) of approximately 65 kDa, and a core protein after chondroitinase ABC digestion of approximately 45 kDa which also contained one to three N-linked oligosaccharides and one O-linked oligosaccharide. The other proteoglycan had an overall size of approximately 100 kDa, two to three keratan sulfate chains of approximately 15 kDa each, and a core protein following keratanase digestion of approximately 51 kDa which included two to three N-linked but no O-linked oligosaccharides. A larger size, a greater overall hydrophobicity (as measured by its interaction with octyl-Sepharose) and an absence of O-linked oligosaccharides argue that this core protein is a distinct gene product from the core protein of the dermatan sulfate proteoglycan.
Midura
R J
RJ
Proteoglycan Chemistry Section, National Institute of Dental Research, Bethesda, Maryland 20892.
Hascall
V C
VC
D000818:Animals; D002642:Chick Embryo; D002807:Chondroitin / Q000031:analogs & derivatives*; D002852:Chromatography, Ion Exchange; D003315:Cornea / Q000378:metabolism*; D003871:Dermatan Sulfate / Q000032:analysis* / Q000096:biosynthesis; D006025:Glycosaminoglycans / Q000032:analysis*; D007632:Keratan Sulfate / Q000032:analysis* / Q000096:biosynthesis; D008970:Molecular Weight
D006025:Glycosaminoglycans; D003871:Dermatan Sulfate; D002807:Chondroitin; D007632:Keratan Sulfate