Serine protease and metallo protease cascade systems involved in pericellular proteolysis.
Cultures of transformed fibroblasts actively involved in extracellular matrix degradation have been examined for initial activation of serine and metallo protease cascade systems. Rous sarcoma virus transformed chick embryo fibroblasts (RSVCEF), in contrast to transformed mammalian cells, produce active, two chain urokinase-type plasminogen activator (tcu-PA). Active tcu-PA is found in serum-free, plasmin-free conditioned medium from RSVCEF cultures as determined by two independent methods, immunoprecipitation and differential DFP sensitivity. RSVCEF cultures synthesize and secrete inactive, single chain uPA (scu-PA) which is converted to tcu-PA in a time dependent manner by a catalytic mechanism that appears to involve a functioning uPA receptor on the surface of intact cells. The enzyme activity responsible for this conversion may represent the initiating catalytic event in the PA/plasminogen serine protease cascade system. A 70 kDa prometalloprotease capable of degrading denatured collagen following its activation also is significantly elevated in RSVCEF cultures over that of normal CEF. Trace amounts of the active 62 kDa form of the metalloprotease (gelatinase) is found in the transformed RSVCEF cultures indicating that these cultures produce a natural activator of the prometalloprotease. Plasmin and/or PA do not appear to be the activator of this enzyme as determined by indirect inhibition assays and direct assays employing purified enzymes. The possible central position of pro PA and the 70 kDa prometalloprotease in an interacting, complex protease cascade system involved in extracellular matrix degradation is discussed.
Quigley
J P
JP
Department of Pathology, SUNY Health Science Center, Stony Brook 11794.
Berkenpas
M B
MB
Aimes
R T
RT
Chen
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D001358:Avian Sarcoma Viruses; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D002642:Chick Embryo; D004789:Enzyme Activation; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology; D018093:Gelatinases; D008322:Mammals; D008565:Membrane Proteins / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism*; D003012:Microbial Collagenase / Q000378:metabolism; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000378:metabolism*; D010434:Pepsin A / Q000378:metabolism; D010960:Plasminogen Activators / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D013045:Species Specificity; D013379:Substrate Specificity; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D016326:Extracellular Matrix Proteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D011506:Proteins; D010960:Plasminogen Activators; D014568:Urokinase-Type Plasminogen Activator; D010434:Pepsin A; D018093:Gelatinases; D008666:Metalloendopeptidases; D003012:Microbial Collagenase