Gene/Protein
Disease
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Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
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Target Concepts:
Gene/Protein
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Query: EC:3.1.6.4 (
chondroitinase
)
2,039
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Retinas were labeled in culture with [3H]glucosamine or [3H]leucine and [35S]sulfate and extracted sequentially with physiologically balanced saline and 4 M guanidine HCl. They were dialyzed into associative conditions (0.5 M NaCl) and chromatographed on agarose columns. Under these conditions, some of the proteoglycans were associated in massive complexes that showed low buoyant densities when centrifuged in CsCl density gradients under dissociative conditions (4 M guanidine HCl). Much of the label in these complexes was in molecules other than proteoglycans. Most of the proteoglycans, however, were included on the agarose columns, where they appeared to be constitutionally of low buoyant density. They resisted attempts to separate potential low buoyant density contaminants from the major proteoglycans by direct CsCl density gradient centrifugation or by the fractionation of saline or 8 M urea extracts on diethylaminoethyl-Sephacel. The diethylaminoethyl-Sephacel fractions were either subjected to CsCl density gradient centrifugation or were chromatographed on Sephacryl S-300, in both cases before and after alkaline cleavage, to confirm the presence of typical O-linked glycosaminoglycans. The medium and balanced salt extracts were enriched in chondroitin sulfate and other sulfated macromolecules, possibly highly sulfated oligosaccharides, that resisted digestion by
chondroitinase
ABC but were electrophoretically less mobile than heparan sulfate.
Guanidine
HCl or urea extracts of the residues were mixtures of high and low density proteoglycans that were enriched in heparan sulfate.
...
PMID:Low buoyant density proteoglycans from saline and dissociative extracts of embryonic chicken retinas. 636 28
Chondroitin sulfate is used extensively as a treatment for osteoarthritis. This study was conducted to evaluate whether chondroitin sulfate could be isolated from chicken keel cartilage in sufficient quantities and of requisite quality to make it a feasible source of chondroitin sulfate. Proteoglycans were extracted from chicken keel cartilage obtained immediately after slaughter by using 3 M MgCl2 at room temperature. The extract was then dialyzed and digested with papain to remove proteins. Glycosaminoglycans were obtained by ethanol precipitation, lyophilized, and characterized by using gel filtration on Sepharose CL-6B columns.
Guanidine
-HCI extraction was also used as a control to investigate the efficiency of extraction using MgCl2. Results showed that, from every gram of wet or non-lyophilized keel cartilage, 32.9 +/- 4.8 mg (dry weight) of glycosaminoglycans could be obtained following MgCl2 extraction. Analyses revealed that 75.5 +/- 4.2% of these glycosaminoglycans were chondroitin sulfate. Chromatographic analyses showed a single symmetrical peak, which could be almost entirely removed by prior digestion with
chondroitinase
ABC, indicating that the material in the peak was in fact chondroitin sulfate. The average molecular weight (also called relative molecular mass, Mr) of the glycosaminoglycans was also estimated (Mr 48,500). Characterization using polyacrylamide or agarose gel electrophoresis showed diffuse bands containing chondroitin sulfate, which could be entirely removed by prior digestion with
chondroitinase
ABC. This study shows that chicken keel cartilage is a readily available source of chondroitin sulfate.
...
PMID:Chicken keel cartilage as a source of chondroitin sulfate. 1216 49