Transient expression of type IV collagenolytic metalloproteinase by human mononuclear phagocytes.
A type IV collagenolytic metalloproteinase secreted by human monocytes/macrophages has been isolated and characterized. Monocytes isolated from peripheral blood and cultured in vitro exhibited a high type IV collagenolytic activity during the first and second day, but such activity declined markedly over subsequent days. Type IV collagenolytic activity was also transiently elaborated by macrophages isolated from (a) bronchioalveolar lavage of patients with pulmonary sarcoidosis, (b) primary human colostrum, and (c) peritoneal lavage of a patient with peritonitis. In contrast, macrophages isolated from the bronchioalveolar lavage of normal individuals, or from noninflammatory peritoneal fluids, failed to exhibit type IV collagenolytic activity. A type IV collagenolytic neutral proteinase was purified from macrophages isolated from inflammatory peritoneal fluid. The proteinase has a mass of 67 kDa on gel electrophoresis and is not altered in its migration under reducing conditions. It produces a characteristic 1/4-3/4 cleavage of type IV collagen, and its activity is abolished by treatment with EDTA but not phenylmethanesulfonyl fluoride. The isoelectric pH of the proteinase is 5.2 as judged by two-dimensional gel electrophoresis. The amino acid composition of the proteinase was notable for a high content of serine, glutamic acid, glycine, and alanine and no detectable hydroxyproline, cysteine, or methionine residues. The carbohydrate content of the proteinase was 11.2%, and galactose was the most abundant monosaccharide (8.7%) released following acid hydrolysis, followed by glucose (1.3%), mannose (1.2%), and trace amounts of fucose and galactosamine. Such a type IV collagenolytic protease may play an important role during the traversal of the vascular basement membrane by extravasating monocytes. The biochemical characteristics and biologic function of the macrophage proteinase may be similar or identical to the type IV collagenolytic proteinase identified in metastatic tumor cells.
D000328:Adult; D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000378:metabolism*; D003126:Colostrum / Q000166:cytology; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D007249:Inflammation; D008264:Macrophages / Q000201:enzymology*; D008297:Male; D008666:Metalloendopeptidases; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D008875:Middle Aged; D009000:Monocytes / Q000201:enzymology; D009363:Neoplasm Proteins / Q000032:analysis; D010529:Peritoneal Cavity / Q000473:pathology; D010538:Peritonitis / Q000473:pathology; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D011247:Pregnancy; D011480:Protease Inhibitors; D011650:Pulmonary Alveoli / Q000473:pathology
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D009363:Neoplasm Proteins; D011480:Protease Inhibitors; D010664:Phenylmethylsulfonyl Fluoride; D003094:Collagen; D004492:Edetic Acid; D010450:Endopeptidases; D008666:Metalloendopeptidases; D003012:Microbial Collagenase