Abstract
Cathepsin B activity, including that of a plasma membrane-associated cathepsin B, has been linked to tumor malignancy. As cathepsin B at the tumor cell surface has been hypothesized to play a role in the focal degradation of basement membrane during the metastatic cascade, we have examined the ability of human tumor cathepsin B to degrade laminin, an adhesive glycoprotein found exclusively in the basement membrane. We report that at pH 6·5 and 7·0 tumor cathepsin B degraded by specific, limited proteolysis both subunits of native laminin. The disappearance of both subunits and the appearance of lower Mr protein bands could be observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Accumulation of degradation products was also observed using gel filtration chromatography and a fluorescamine assay. The proteolysis of laminin by tumor cathepsin B could be inhibited by an active site titrant for cysteine proteinases or stefin B, an endogenous low-Mr cysteine proteinase inhibitor.
Similar content being viewed by others
References
Anastasi, A., Brown, M. A., Kembhavi, A. A., Nicklin, M. J. H., Sayers, C. A., Sunter, D. C., andBarrett, A. J., 1983, Cystatin, a protein inhibitor of cysteine proteinases.Biochemical Journal,211, 129–138.
Barrett, A. J., 1977, Cathepsin B and other thiol proteinases.Proteinases in Mammalian Cells and Tissues, edited by A. J. Barrett (Amsterdam: Elsevier), pp. 181–208.
Barrett, A. J., andKirschke, H., 1980, Cathepsin B, cathepsin H and cathepsin L.Methods in Enzymology,80, 535–563.
Crissman, J. D., Hatfield, J. S., andHonn, K. V., 1986, Arrest and extravasation of circulating murine tumor cells in the lung.Laboratory Investigation,54, 14 (abstract).
Crissman, J. D., Hatfield, J. S., Schaldenbrand, M., Sloane, B. F., andHonn, K. V., 1986, Arrest and extravasation of B16 amelanotic melanoma in murine lungs.Laboratory Investigation,53, 470–478.
Green, G. D. J., Kembhavi, A. A., Davies, M. E., andBarrett, A. J., 1984, Cystatin-like cysteine proteinase inhibitors from human liver.Biochemical Journal,218, 939–946.
Laemmli, U.K., 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature (London),227, 680–685.
Liotta, L. A., Rao, N. C., andBarsky, S. H., 1983, Tumor invasion and the extracellular matrix.Laboratory Investigation,49, 636–649.
Mort, J. S., Leduc, M. S., andRecklies, A. D., 1983, Characterization of a latent cysteine proteinase from ascitic fluid as a high molecular weight form of cathepsin B.Biochimica et Biophysica Acta,755, 369–375.
Quigley, J., Cramer, A., Fairbairn, S., Gilbert, R., Lacovara, J., Ojakian, G., andSchwimmer, R., 1985, Interaction of malignant cells with substrata: adhesion, degradation and migration.Mechanisms of Cancer Metastasis: Potential Therapeutic Implications, edited by K. V. Honn, W. E. Powers and B. F. Sloane (Boston: Martinus Nijhoff), pp. 309–339.
Rao, N. C., Margulies, I.M. K., Goldfarb, R. H., Madri, J. A., Woodely, D.T.,andLiotta, L. A., 1982, Differential proteolytic susceptibility of laminin alpha and beta subunits.Archives of Biochemistry and Biophysics,219, 65–70.
Rich, D. H., Brown, M. A., andBarrett, A. J., 1986, Purification of cathepsin B by a new form of affinity chromotography.Biochemical Journal,235, 731–734.
Ryan, R. E., Crissman, J. D., Honn, K. V., andSloane, B. F., 1985, Cathepsin B-like activity in viable tumor cells isolated from rodent tumors.Cancer Research,45, 3636–3641.
Sloane, B. F., andBird, J. W. C., 1977, Effect of ovarian hormones on lysosomal acid hydrolase activities in rat myometrium.American Journal of Physiology,223, E423-E431.
Sloane, B. F., Lah, T. T., Day, N. A., Rozhin, J., Bando, Y., andHonn, K. V., 1986, Tumor cysteine proteinases and their inhibitors.Cysteine Proteinases and Their Inhibitors, edited by V. Turk (Berlin: Walter de Gruyter), pp. 729–749.
Sloane, B. F., Rozhin, J., Johnson, K., Taylor, H., Crissman, J. D., andHonn, K. V., 1986, Cathepsin B: association with plasma membrane in metastatic tumors.Proceedings of the National Academy of Sciences of the U.S.A.,83, 2483–2487.
Takahashi, T., Dehdarani, A. H., Yonezawa, S., andTang, J., 1986, Comparative studies of two cathepsin B isozymes from porcine spleen.Journal of Biological Chemistry,261, 9315–9381.
Terranova, V. P., Liotta, L. A., Russo, R. G., andMartin, G. R., 1982, Role of laminin in the attachment and metastasis of murine tumor cells.Cancer Research,42, 2265–2269.
Timpl, R., Rhoe, H., Robey, P. G., Rennard, S. I., Foidart, J. M., andMartin, G. R., 1979, Laminin—a glycoprotein from basement membranes.Journal of Biological Chemistry,254, 9933–9937.
Willenbrock, F., andBrocklehurst, K., 1985, Preparation of cathepsins B and H by covalent chromatography and characterization of their catalytic sites by reaction with a thiol specific two-protonic-state reactivity probe.Biochemical Journal,227, 511–519.
Yamada, K. M., 1983, Cell surface interactions with extracellular materials.Annual Review of Biochemistry,52, 761–799.
Author information
Authors and Affiliations
Additional information
On leave from the Jozef Stefan Institute, Ljubljana, Yugoslavia.
Rights and permissions
About this article
Cite this article
Lah, T.T., Buck, M.R., Honn, K.V. et al. Degradation of laminin by human tumor cathepsin B. Clin Exp Metast 7, 461–468 (1989). https://doi.org/10.1007/BF01753666
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01753666