title: Partial purification and characterization of serum protease from tumor-bearing rats which cleaves type IV collagen.
abstract: Activity of neutral protease was increased in sera of rats bearing ascites hepatoma AH109A compared to those of normal rats. The protease was isolated from serum protein and partially purified approximately 1,150 times in specific activity after sequential column chromatography of hemoglobin affinity, lysine-Sepharose, Ultrogel AcA34 and TSK-gel G2000SW in that order. The protease fraction still seemed to contain at least two kinds of proteases, serine and cysteine protease. It had a molecular weight of 18-21 kilodaltons with broad optimal pH range of 7.0-9.0, maximum at 8.0. Intradermal injection of the crude preparation of the neutral protease fraction induced extravascular emigration of circulating tumor cells in vivo. Moreover, partially purified protease degraded pepsin-treated chains of bovine glomerular type IV collagen in vitro, but such an in vitro action of the protease was inhibited by an addition of soybean trypsin inhibitor or mercuric chloride. It failed to cleave salt-extracted rat skin type I collagen under the same digestive conditions for bovine type IV collagen. The serum neutral proteases of tumor-bearing host may play some cooperative roles during extravascular emigration of tumor cells by destruction of vascular basement membrane.
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000032:analysis; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003094:Collagen / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D009362:Neoplasm Metastasis; D010447:Peptide Hydrolases / Q000302:isolation & purification*; D011480:Protease Inhibitors / Q000378:metabolism; D051381:Rats; D012867:Skin / Q000378:metabolism
chemical_list: D001798:Blood Proteins; D011480:Protease Inhibitors; D003094:Collagen; D010447:Peptide Hydrolases