Structural studies of two populations of keratan sulphate chains from mature bovine articular cartilage.
Two discrete peptido-keratan sulphate fragments were isolated via chondroitinase ABC and trypsin digestion of a proteoglycan aggregate fraction prepared from bovine femoral head cartilage (six year old animals). The larger fragments (K(av) = 0.07, CL-6B) contained peptides substituted with several keratan sulphate (KS) chains from the KS-rich region of the proteoglycan and the smaller fragments (K(av) = 0.5, CL-6B) contained peptides with, perhaps, only one KS chain and the stubs of post-chondroitinase-treated chondroitin sulphate chains. The two peptido-KS samples and the KS chains derived from these by alkaline borohydride reduction were characterised by 13C-NMR spectroscopy. The two populations of KS chains were also examined by chromatography (Sephadex G-75), and keratanase digestion followed by chromatography on Bio-Gel P-10. From the results it was concluded that the KS chains from the two major trypsin-derived peptido-KS fragments had similar sulphation levels, distributions of hydrodynamic sizes and susceptibilities to keratanase.
Thornton
D J
DJ
Division of Biological Sciences, University of Lancaster, U.K.
Morris
H G
HG
Cockin
G H
GH
Huckerby
T N
TN
Nieduszynski
I A
IA
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002358:Cartilage, Articular / Q000737:chemistry*; D002417:Cattle; D002810:Chondroitinases and Chondroitin Lyases; D002850:Chromatography, Gel; D010450:Endopeptidases; D007202:Indicators and Reagents; D007632:Keratan Sulfate / Q000737:chemistry* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D010446:Peptide Fragments / Q000302:isolation & purification; D014357:Trypsin
D002241:Carbohydrates; D007202:Indicators and Reagents; D010446:Peptide Fragments; D007632:Keratan Sulfate; D010450:Endopeptidases; D014357:Trypsin; D002810:Chondroitinases and Chondroitin Lyases