Processing and intracellular transport of cathepsin G and neutrophil elastase in the leukemic myeloid cell line U-937-modulation by brefeldin A, ammonium chloride, and monensin.
The effects of brefeldin A, monensin, and the weak base NH4Cl on the biosynthesis and processing of cathepsin G and neutrophil elastase of myeloid cells were investigated. Monoblast-like U-937 cells were biosynthetically labeled with [35S]methionine, followed by subcellular fractionation, immunoprecipitation, and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Brefeldin A inhibited proteolytic processing, intracellular transport, and secretion. The effects were reversible inasmuch as removal of brefeldin A resulted in a normal pattern of processing and transfer to high-density organelles, corresponding to lysosomes, and restitution of constitutive secretion of precursor forms. Both cathepsin G and neutrophil elastase acquired resistance to endoglycosidase-H, suggesting that conversion to complex oligosaccharide side chains also occurs in the presence of brefeldin A. Monensin and NH4Cl inhibited final proteolytic processing, indicating either that acidification is necessary for directing cathepsin G and neutrophil elastase to lysosomal-like organelles or that the protease(s) responsible for processing requires an acid pH. We conclude that pH-dependent proteolytic processing of cathepsin G and neutrophil elastase occurs in post-Golgi structures and that transfer to lysosomes or an immediately prelysosomal compartment is mandatory for complete processing.
Lindmark
A
A
Research Department, Lund Hospital, Sweden.
Gullberg
U
U
Olsson
I
I
D000643:Ammonium Chloride / Q000494:pharmacology*; D000935:Antifungal Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D020126:Brefeldin A; D056649:Cathepsin G; D002403:Cathepsins / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007951:Leukemia, Myeloid / Q000201:enzymology*; D019272:Leukocyte Elastase; D008985:Monensin / Q000494:pharmacology*; D010196:Pancreatic Elastase / Q000378:metabolism*; D009195:Peroxidase / Q000378:metabolism; D012697:Serine Endopeptidases; D014407:Tumor Cells, Cultured
D000935:Antifungal Agents; D003517:Cyclopentanes; D000643:Ammonium Chloride; D020126:Brefeldin A; D008985:Monensin; D009195:Peroxidase; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase