Isolation and characterization of a metalloproteinase secreted by rat glioma cells in serum-free culture.

Abstract

The isolation of a metalloproteinase secreted by a rat glioma cell line (BT5C) in serum-free media is described. After affinity purification, the activity was present as a double band with Mr 86000 and 76000 both of which required CaCl2 for activity. The enzyme was able to degrade gelatin but not casein. It was unable to degrade native types I, III, IV and V collagens but their denatured counterparts were degraded. Using a radiolabel release assay the enzyme was inhibited by EDTA, 1:10 phenanthroline and TIMP confirming that it belongs to the family of metalloproteinases. Its activity was not affected by either serine or cysteine protease inhibitors. The proteinase was activated by APMA but was unaffected by trypsin treatment.

• Authors: G J Rucklidge, M Lund-Johansen, G Milne, R Bjerkvig
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 172 Issue 2 Pg. 544-50 (Oct 30 1990) ISSN: 0006-291X [Print] United States
• PMID: 2241953 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Culture Media
• Protease Inhibitors
• Metalloendopeptidases

Topics

• Animals
• Cell Line
• Chromatography, Affinity
• Culture Media
• Electrophoresis, Polyacrylamide Gel
• Enzyme Activation
• Glioma
• Kinetics
• Metalloendopeptidases (isolation & purification, metabolism)
• Protease Inhibitors (pharmacology)
• Rats