alpha-chymotrypsin and Fibrosarcoma

Matrix metalloproteinase 9 (MMP-9) has been purified as an inactive zymogen of M(r) 92,000 (proMMP-9) from the culture medium of HT 1080 human fibrosarcoma cells. The NH2-terminal sequence of proMMP-9 is Ala-Pro-Arg-Gln-Arg-Gln-Ser-Thr-Leu-Val-Leu-Phe-Pro, which is identical to that of the 92-kDa type IV collagenase/gelatinase. The zymogen can be activated by 4-aminophenylmercuric acetate, yielding an intermediate form of M(r) 83,000 and an active species of M(r) 67,000, the second of which has a new NH2 terminus of Met-Arg-Thr-Pro-Arg-(Cys)-Gly-Val-Pro-Asp-Leu-Gly-Arg-Phe-Gln-Thr- Phe-Glu. Immunoblot analyses demonstrate that this activation process is achieved by sequential processing of both NH2- and COOH-terminal peptides. TIMP-1 complexed with proMMP-9 inhibits the conversion of the intermediate form to the active species of M(r) 67,000. The proenzyme is fully activated by cathepsin G, trypsin, alpha-chymotrypsin, and MMP-3 (stromelysin 1) but not by plasmin, leukocyte elastase, plasma kallikrein, thrombin, or MMP-1 (tissue collagenase). During the activation by MMP-3, proMMP-9 is converted to an active species of M(r) 64,000 that lacks both NH2- and COOH-terminal peptides. In addition, HOCl partially activates the zymogen by reacting with an intermediate species of M(r) 83,000. The enzyme degrades type I gelatin rapidly and also cleaves native collagens including alpha 2 chain of type I collagen, collagen types III, IV, and V at undenaturing temperatures. These results indicate that MMP-9 has different activation mechanisms and substrate specificity from those of MMP-2 (72-kDa gelatinase/type IV collagenase).

Topics: Amino Acid Sequence; Blotting, Western; Cathepsin G; Cathepsins; Chymotrypsin; Collagenases; Electrophoresis, Polyacrylamide Gel; Enzyme Activation; Enzyme Precursors; Fibrosarcoma; Glycoproteins; Humans; Kinetics; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Metalloendopeptidases; Molecular Sequence Data; Phenylmercuric Acetate; Serine Endopeptidases; Substrate Specificity; Tissue Inhibitor of Metalloproteinases; Trypsin; Tumor Cells, Cultured

Forty-one cases of dermatofibrosarcoma protuberans are presented. The clinical features and histopathological appearances are described. Immunohistochemical staining of thirteen cases with antisera to lysozyme, alpha 1-antichymotrypsin and S-100 protein has provided no evidence to support either a histiocytic or neuroectodermal origin for these tumours. In reviewing the literature, the histogenetic origin, differential diagnosis and malignant potential of dermatofibrosarcoma protuberans are discussed.

Topics: Adolescent; Adult; Aged; alpha 1-Antichymotrypsin; Chymotrypsin; Female; Fibrosarcoma; Humans; Male; Middle Aged; Muramidase; S100 Proteins; Skin Neoplasms

Media conditioned by tumor cells were studied for the presence of factor(s) that increase the rate of random migration (chemokinesis) of Corynebacterium parvum-activated macrophages. A capillary tube assay was developed and utilized to expediently monitor the chemokinetic activity of macrophages incubated in whole and fractionated media. Media conditioned by six different syngeneic and allogeneic mouse tumor cell lines demonstrated significantly higher chemokinetic activity than unconditioned or normal fibroblast conditioned media. The chemokinetically active component of the Lewis Lung conditioned media was found to be a trypsin sensitive, heat stable, high molecular weight (300,000-480,000 dalton range) factor that had no chemotactic (directional migration) activity. Pyran-activated macrophages also responded chemokinetically to the Lewis Lung factor while oyster glycogen and thioglycolate-elicited macrophages did not. The similarity and differences between the physical properties of the chemokinetic factor, other migration stimulating factors, and tumor-associated proteins are discussed.

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Line; Cell Movement; Chemotactic Factors; Chemotaxis; Chymotrypsin; Fibrosarcoma; Lung Neoplasms; Macrophages; Male; Mammary Neoplasms, Experimental; Melanoma; Mice; Mice, Inbred C57BL; Neoplasms, Experimental; Trypsin

A variant subpopulation of C57BL/6 mouse fibrosarcoma cells that had very low tumorigenic potential was isolated from a highly tumorigenic parent fibrosarcoma cell culture. The adhesive characteristics of parent cells and variant cells were compared. The low-tumorigenic variant cells were released from the surfaces of plastic dishes, from protein-coated dishes, or from monolayers of fibroblasts or endothelial cells by protease treatment much more readily than were the parent cells. There was no difference between the variant cells and the parent cells in EDTA sensitivity or sensitivity to mechanical agitation under the conditions used. Also, no difference existed between the variant cells and the parent cells in rates of attachment to the surfaces of plastic dishes or to monolayers of endothelial cells. The variant cells were characterized by high levels of chymotrypsin-like esterase activity (two to three times increased over parent cell levels), but there was only a slight difference between the variant cells and the parent cells in caseinolytic or fibrinolytic activity.

Topics: Animals; Caseins; Cell Adhesion; Cell Separation; Cells, Cultured; Chymotrypsin; Edetic Acid; Fibrin; Fibrosarcoma; Mice; Neoplasms, Experimental; Peptide Hydrolases

Topics: Animals; Cell Movement; Chymotrypsin; Clone Cells; Esterases; Female; Fibrosarcoma; Glucuronidase; Glycoside Hydrolases; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasm Transplantation; Sarcoma, Experimental

Topics: Animals; Antigens, Neoplasm; Bacterial Infections; Benzopyrenes; Cell Migration Inhibition; Chromatography, Gel; Chymotrypsin; Fibrosarcoma; Intradermal Tests; Leukocytes; Lymphocytes; Male; Molecular Weight; Rats; Rats, Inbred WF; Spleen; T-Lymphocytes; Thoracic Duct

Topics: Animals; Antigens, Neoplasm; Benzene Derivatives; Boron Compounds; Bromelains; Chelating Agents; Chymotrypsin; Cricetinae; Edetic Acid; Endopeptidases; Enzymes; Fibrosarcoma; Histocompatibility Antigens; Hyaluronoglucosaminidase; Injections, Subcutaneous; Microbial Collagenase; Neoplasm Transplantation; Pancreatin; Papain; Sarcoma, Experimental; Simian virus 40; Transplantation, Homologous; Vaccines