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PMID: 23763 Published · ppublish English Journal Article

Proteoglycan-degrading enzymes. A radiochemical assay method and the detection of a new enzyme cathepsin F.

The Biochemical journal ·Vol. 167 ·No. 3 ·1977-12-01 ·Pages 775-85

Dingle JT, Blow AM, Barrett AJ, Martin PE

Abstract

1. Polyacrylamide beads containing entrapped 35S-labelled proteoglycan molecules have been prepared. 2. The measurement of release of radioactivity provides an extremely sensitive assay for proteoglycan-degrading enzymes, including proteinases and hyaluronidase. 3. The amount of label released is a logarithmic function of enzyme concentration or time of incubation. Experiments were made in an attempt to explain this. 4. Assays were made by the new method at several pH values, and with the inclusion of inhibitors to identify the proteoglycan-degrading enzymes of rabbit ear cartilage. 5. A previously undescribed proteinase active against proteoglycan at pH4.5 but unaffected by pepstatin, was discovered. The enzyme was named cathepsin F, and was partially purified and characterized; it was detected in human articular cartilage.

MeSH Terms
Acrylamides Animals Cartilage/enzymology Cathepsins/analysis,antagonists & inhibitors,isolation & purification Chromatography, Gel Humans Hydrogen-Ion Concentration Immunoglobulin G Pepstatins/pharmacology Proteoglycans Rabbits Sepharose Subcellular Fractions/enzymology Sulfur Radioisotopes Viscosity
Chemicals
Acrylamides Immunoglobulin G Pepstatins Proteoglycans Sulfur Radioisotopes Sepharose Cathepsins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Dingle J T
Blow A M
Barrett A J
Martin P E
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25 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1670775
Published
1977-12-01
Pages
775-85
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1183726
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1670775 PMC Full Text View journal details
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