The processing and intracellular transport of myeloperoxidase. Modulation by lysosomotropic agents and monensin.
Myeloperoxidase, stored in azurophil granules of neutrophils, is synthesized in promyelocytes as a larger molecular weight precursor, which is processed to yield a transient Mr 82 000 intermediate and mature polypeptides with molecular weights of 62 000 and 12 000. We have tried to define subcellular sites for processing using metabolic labelling of the promyelocytic leukemia cell line HL-60 in combination with subcellular fractionation on a Percoll gradient. A reasonable separation was achieved between azurophil granules, Golgi elements and endoplasmic reticulum. The finding of almost exclusively fully processed myeloperoxidase in granules and a mixture of unprocessed and processed polypeptide in fractions enriched in Golgi elements suggests that processing occurred mainly in pregranular structures. Monensin, which exchanges protons for Na+, and the base chloroquine blocked processing probably by inhibition of transport through the Golgi apparatus. However, the lysosomotropic NH4+ cation did not inhibit processing or transport indicating that processing is not necessarily influenced by pH-dependent mechanisms. Results from digestion with endoglycosidase H, incubation with tunicamycin and metabolic labelling with [3H]mannose indicated that myeloperoxidase contained high mannose oligosaccharide side chains. Also [32P]phosphate incorporated into Mr 90 000 and Mr 62 000 myeloperoxidase was susceptible to endoglycosidase H indicating that oligosaccharide side chains are modified by phosphorylation as in lysosomal enzymes. Thus, even if myeloperoxidase contained mannose 6-phosphate residues, these may not necessarily be involved in directing transport to the azurophil granules.
D000643:Ammonium Chloride / Q000494:pharmacology; D001693:Biological Transport, Active / Q000187:drug effects; D001826:Body Fluids / Q000201:enzymology*; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D005663:Furans / Q000494:pharmacology*; D006801:Humans; D007424:Intracellular Fluid / Q000201:enzymology*; D008247:Lysosomes / Q000187:drug effects*; D008970:Molecular Weight; D008985:Monensin / Q000494:pharmacology*; D009844:Oligosaccharides / Q000378:metabolism; D009195:Peroxidase / Q000032:analysis / Q000378:metabolism*; D010766:Phosphorylation
D005663:Furans; D009844:Oligosaccharides; D000643:Ammonium Chloride; D002738:Chloroquine; D008985:Monensin; D009195:Peroxidase