TY - JOUR
T1 - Isolation and characterization of a metalloproteinase secreted by rat glioma cells in serum-free culture
AU - Rucklidge, Garry J.
AU - Lund-Johansen, Morten
AU - Milne, George
AU - Bjerkvig, Rolf
N1 - Funding Information:
ICRETT grant from WCC.
Funding Information:
’ Supported by The Norwegian Cancer Society. G.J.R acknowledges the receipt of an
PY - 1990/10/30
Y1 - 1990/10/30
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0025153043&partnerID=8YFLogxK
U2 - 10.1016/0006-291X(90)90707-T
DO - 10.1016/0006-291X(90)90707-T
M3 - Article
C2 - 2241953
AN - SCOPUS:0025153043
SN - 0006-291X
VL - 172
SP - 544
EP - 550
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -