title: Cathepsin B: association with plasma membrane in metastatic tumors.
abstract: The subcellular localization of cathepsin B activity (EC 3.4.22.1) in three murine melanomas of increasing metastatic potential (Cloudman less than B16-F1 less than B16 amelanotic) was determined. Cathepsin B activity was localized in the heavy mitochondrial fraction of normal murine liver but in the light mitochondrial fraction of the metastatic melanomas; the localization of three other lysosomal hydrolases did not shift. Further purification of the light mitochondrial fraction into L-1 (density = 1.045 g/ml) and L-2 (density = 1.07 g/ml) fractions was achieved on a 30% iso-osmotic Percoll gradient. The L-1 fraction of liver and melanomas contained Na+, K+-ATPase activity; the L-2 fraction of liver contained four lysosomal hydrolase (cathepsins B and H, N-acetyl-beta-glucosaminidase, and beta-glucuronidase) and glucose-6-phosphatase activities. Ultrastructural examination revealed that the L-1 fraction consisted of membrane vesicles and the L-2 fraction of secondary lysosomes. In the B16 melanomas cathepsin B and N-acetyl-beta-glucosaminidase activities were found in both L-1 and L-2 fractions. Specific activities of the two enzymes in the plasma membrane (L-1) fractions increased in correspondence with metastatic potential. Cathepsin H and beta-glucuronidase activities were not localized in the plasma membrane fractions of the B16 melanomas. Localization of hydrolytic enzymes in the plasma membrane of metastatic tumor cells could result in focal dissolution of the extracellular matrix and thereby invasion and metastasis.
authors.forename: B F
authors.initials: BF
authors.lastname: Sloane
authors.forename: J
authors.initials: J
authors.lastname: Rozhin
authors.forename: K
authors.initials: K
authors.lastname: Johnson
authors.forename: H
authors.initials: H
authors.lastname: Taylor
authors.forename: J D
authors.initials: JD
authors.lastname: Crissman
authors.forename: K V
authors.initials: KV
authors.lastname: Honn
mesh_terms: D000118:Acetylglucosaminidase / Q000378:metabolism; D000818:Animals; D002401:Cathepsin B; D002403:Cathepsins / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D008099:Liver / Q000201:enzymology / Q000648:ultrastructure; D008175:Lung Neoplasms / Q000556:secondary / Q000648:ultrastructure; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D008545:Melanoma / Q000201:enzymology / Q000473:pathology / Q000648:ultrastructure; D051379:Mice; D009362:Neoplasm Metastasis; D009374:Neoplasms, Experimental / Q000201:enzymology* / Q000473:pathology / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D000118:Acetylglucosaminidase; D002403:Cathepsins; D002401:Cathepsin B