A growth-regulated protease activity that is inhibited by the anticarcinogenic Bowman-Birk protease inhibitor.
The Bowman-Birk protease inhibitor (BBI) has been shown to be an effective suppressor of carcinogenesis in vivo and in vitro. To elucidate the mechanism(s) by which BBI suppresses carcinogenesis, we believe it will be necessary to identify and characterize the target enzymes that specifically interact with the BBI. We have shown previously that several cellular proteins in C3H/10T1/2 mouse embryo fibroblast cells specifically bind to a BBI affinity resin. In the current report, we demonstrate that one of these proteins has proteolytic activity as judged by its ability to degrade gelatin. The enzyme has a mass of 45 kDa and subcellular fractionation experiments demonstrate that this enzyme is located in the cytosol. Furthermore, the proteolytic activity was inhibited by diisopropylfluorophosphate but was not affected by EDTA, indicating that this enzyme is a serine protease. Higher levels of protease activity were found in logarithmic-phase C3H/10T1/2 cells compared with nondividing (confluent) cells, suggesting that this protease activity is growth regulated. Similar levels of this activity were present in nontransformed and in radiation-transformed C3H/10T1/2 cells. Treatment of nontransformed C3H/10T1/2 cells with phorbol 12-myristate 13-acetate increased the specific activity of this protease 5- to 10-fold. Our results suggest that this protease is a target enzyme of the BBI in these cells.
Billings
P C
PC
Department of Radiation Oncology, University of Pennsylvania School of Medicine, Philadelphia 19104.
Habres
J M
JM
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D002455:Cell Division; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D066298:In Vitro Techniques; D051379:Mice; D008970:Molecular Weight; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013347:Subcellular Fractions / Q000201:enzymology; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000378:metabolism / Q000494:pharmacology*
D000970:Antineoplastic Agents; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D012697:Serine Endopeptidases