title: Enrichment of collagen and gelatin degrading activities in the plasma membranes of human cancer cells.
abstract: Interactions between connective tissue substrates and proteinases localized to the surface of cancer cells are implicated in cancer invasion. In this report we have compared the enrichment of collagen and gelatin degrading activities and cysteine proteinase(s) in well-characterized (enzyme markers and electron microscopy) subcellular membrane fractions isolated from human small cell lung cancer lines (NCI-H69 and NCI-H82) and the RWP-1 pancreatic cancer line. With each cell line collagenolytic, gelatinolytic, and cysteine proteinase activities were enriched 5- to 128-fold in the plasma membrane fractions with differences noted between microvilli versus smooth membrane profiles. Incubation of tumor plasma membranes with methyl-3H-labeled collagen resulted in extensive degradation of the gamma, beta, alpha 1, and alpha 2 chains, suggesting the combined action of metalloproteinases. Treatment of tumor plasma membranes with the chaotropic agent, 2 M KCl, did not diminish membrane collagen- or gelatin-degrading activity, but extensively leached out the cysteine proteinase, suggesting that the latter enzyme is not an integral membrane protein. Enzyme inhibitors specific for metalloproteinases and cysteine proteinase were used to corroborate enzymatic classification. In conclusion, we have demonstrated variations in the localization of proteinases in the plasma membrane domains of different human cancer cells.
mesh_terms: D002462:Cell Membrane / Q000201:enzymology*; D003094:Collagen / Q000378:metabolism*; D003546:Cysteine Endopeptidases; D010450:Endopeptidases / Q000032:analysis; D005780:Gelatin / Q000378:metabolism*; D006801:Humans; D008666:Metalloendopeptidases; D009369:Neoplasms / Q000201:enzymology* / Q000648:ultrastructure; D011189:Potassium Chloride / Q000494:pharmacology
chemical_list: D011189:Potassium Chloride; D005780:Gelatin; D003094:Collagen; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D008666:Metalloendopeptidases