Chondroitin sulfate is involved in lysosomal transport of lysozyme in U937 cells.
Human promonocytes U937 synthesize lysozyme and retain approximately one third of it within lysosomes. Lysozyme is not glycosylated; thus, it cannot be subject to mannose-6-phosphate-dependent targeting to lysosomes. It is a basic protein with a pI of 10.5 and is known to interact with negatively charged macromolecules like proteoglycans. Therefore, we examined whether the latter are involved in the lysosomal targeting of lysozyme in U937 cells. We partially diminished the electronegative charge of newly synthesized proteoglycans by inhibiting their sulfation with chlorate. This increased the rate of secretion of lysozyme. Upon treatment of U937 cells with phorbol esters, the rate of secretion of lysozyme was increased to more than 90%. This coincided with an almost complete redistribution of a [(35)S]sulfate bearing proteoglycan to the secretory pathway. After a brief pulse with [(35)S]sulfate in the control, 80% of the [(35)S]sulfate-bearing proteoglycan was retained within the cells, whereas in the treated cells this proportion was decreased to 13%. The secreted proteoglycan was sensitive to chondroitinase ABC and bound to immobilized lysozyme. This interaction was disrupted by 50-300 mM NaCl. The intracellularly retained proteoglycan was degraded with a half-life of 50-60 minutes and seemed to be directed to lysosomes because in the presence of NH(4)Cl the degradation was strongly inhibited. Our results suggest that the proteoglycan is involved in lysosomal targeting of lysozyme in U937 cells.
Lemansky
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P
Philipps-Universität Marburg, Institut für Physiologische Chemie, Karl-von-Frisch-Strasse 1, Germany.
Hasilik
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A
D000643:Ammonium Chloride / Q000494:pharmacology; D019765:Chondroitin ABC Lyase; D002809:Chondroitin Sulfates / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D008360:Mannosephosphates / Q000378:metabolism; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*; D010472:Perchlorates / Q000494:pharmacology; D011485:Protein Binding; D021381:Protein Transport; D011509:Proteoglycans / Q000378:metabolism; D017670:Sodium Compounds / Q000494:pharmacology; D013431:Sulfates / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D020298:U937 Cells
D004800:Enzymes, Immobilized; D008360:Mannosephosphates; D010472:Perchlorates; D011509:Proteoglycans; D017670:Sodium Compounds; D013431:Sulfates; D013462:Sulfur Radioisotopes; D000643:Ammonium Chloride; C027693:mannose-6-phosphate; D002809:Chondroitin Sulfates; C031068:sodium perchlorate; D009113:Muramidase; D019765:Chondroitin ABC Lyase; D013755:Tetradecanoylphorbol Acetate