Identification of specific binding sites for keratan sulphate proteoglycans and chondroitin-dermatan sulphate proteoglycans on collagen fibrils in cornea by the use of cupromeronic blue in 'critical-electrolyte-concentration' techniques.
Proteoglycans (PGs) in bovine corneal stroma were stained with Cupromeronic Blue in 'critical-electrolyte-concentration' (CEC) methods for electron microscopy, and were located vis-à-vis collagen fibril a-e banding patterns. Keratanase and chondroitin ABC lyase digestion showed that a + c-band- and d + e-band-associated PGs were keratan sulphate-rich and chondroitin (dermatan) sulphate-rich respectively. The CEC pattern proved that the keratan sulphate PGs at the a and c bands differed. Comparison of their CECs with their behaviour on anion-exchange chromatography confirmed previous (indirect) attempts at identification [Scott & Haigh (1985) Biosci. Rep. 5, 765-774]. Similar arguments were applied to the dermatan sulphate PGs at the d and e bands. These results strongly support the one-PG-one-binding-site hypothesis [e.g. Scott (1988) Biochem. J. 252, 313-323]. Remarkable inter-species variations in the keratan sulphate PG patterns contrast with the relatively constant picture of dermatan sulphate PG-collagen fibril interactions.
Scott
J E
JE
Chemical Morphology, Cell and Structural Biology, University of Manchester, U.K.
Haigh
M
M
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002807:Chondroitin / Q000031:analogs & derivatives*; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism*; D004396:Coloring Agents; D003315:Cornea / Q000378:metabolism* / Q000648:ultrastructure; D003871:Dermatan Sulfate / Q000378:metabolism*; D004573:Electrolytes; D006025:Glycosaminoglycans / Q000378:metabolism*; D007202:Indicators and Reagents; D007211:Indoles; D007632:Keratan Sulfate / Q000378:metabolism*; D000071234:Lumican; D008854:Microscopy, Electron; D009942:Organometallic Compounds; D011509:Proteoglycans / Q000378:metabolism*; D011817:Rabbits
D011508:Chondroitin Sulfate Proteoglycans; D004396:Coloring Agents; D004573:Electrolytes; D006025:Glycosaminoglycans; D007202:Indicators and Reagents; D007211:Indoles; D000071234:Lumican; D009942:Organometallic Compounds; D011509:Proteoglycans; C032649:dermatan sulfate proteoglycan; D003871:Dermatan Sulfate; C015445:copper phthalocyanine; D002807:Chondroitin; D003094:Collagen; D007632:Keratan Sulfate