title: A 170-kDa membrane-bound protease is associated with the expression of invasiveness by human malignant melanoma cells.
abstract: Malignant spreading of cancer cells requires cell surface proteases that cleave the crosslinked collagenous matrix of connective tissues. From correlating the morphologically defined invasiveness of tumor cells with the presence of specific membrane-associated proteases, we have identified a malignant human melanoma cell line, LOX, that invades crosslinked gelatin films in vitro and contains uniquely a neutral 170-kDa gelatinase in the cell membrane. A similar gelatinase was found in membranes recovered from culture media conditioned with LOX. The 170-kDa gelatinase is a wheat germ agglutinin-binding protein. The proteolytic activity is maximal at neutral pH, enhanced by EDTA and dithiothreitol, inhibited by the cysteine protease inhibitors N-ethylmaleimide, HgCl2, and phenylmethylsulfonyl fluoride, and can bind to an organomercurial adsorbent, suggesting that it is a neutral sulfhydryl-sensitive protease. This 170-kDa gelatinase of LOX cells was not found in a control melanoma cell line, SK-MEL28, or in 32 other tumor cell lines that did not show extracellular gelatin degradation. Thus, we have identified a large membrane-bound protease that may be a specific marker molecule for melanoma cell invasiveness.
authors.affiliation: Department of Anatomy and Cell Biology, Georgetown University School of Medicine, Washington, DC 20007.
authors.forename: A
authors.initials: A
authors.lastname: Aoyama
authors.forename: W T
authors.initials: WT
authors.lastname: Chen
mesh_terms: D002458:Cell Fractionation; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D018093:Gelatinases; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D008545:Melanoma / Q000201:enzymology* / Q000473:pathology; D008970:Molecular Weight; D009361:Neoplasm Invasiveness; D010434:Pepsin A / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D010450:Endopeptidases; D010434:Pepsin A; D018093:Gelatinases