Studies on the extraction of different proteoglycan populations in bovine articular cartilage.
Large proteoglycan monomers and small dermatan sulfate proteoglycans were extracted from explants of bovine articular cartilage with increasing (0-4 M) concentrations of guanidinium chloride (GuHCl). The first extractions were followed by a second extraction with 4 M GuHCl. The amount of proteoglycans extracted in the first buffer depended on the GuHCl concentration. At low concentrations of GuHCl, a relatively high amount of small proteoglycans was obtained. Fifty percent of the small proteoglycans was extracted in buffer with 0.85 M GuHCl, while 2.0-2.2 M GuHCl was needed to extract half of the large proteoglycans. Immediately after synthesis, 35S-labeled large proteoglycans were extracted much easier (50% at 1.4 M GuHCl), and those extracted at low concentrations of GuHCl were less capable of aggregation with hyaluronic acid. After 7 days of 'chase' these differences between endogenous and 35S-labeled proteoglycans had disappeared.
Brand
H S
HS
Jan van Breemen Instituut, Amsterdam.
Korver
G H
GH
van de Stadt
R J
RJ
van Kampen
G P
GP
van der Korst
J K
JK
D000818:Animals; D002021:Buffers; D002358:Cartilage, Articular / Q000737:chemistry*; D002417:Cattle; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011509:Proteoglycans / Q000302:isolation & purification*
D002021:Buffers; D011509:Proteoglycans