Degradation of endocytosed dermatan sulfate proteoglycan in human fibroblasts.
Endocytosis and subsequent degradation of iduronic acid-rich small dermatan sulfate proteoglycan from fibroblast secretions were studied in human fibroblasts. Upon endocytosis of [3H]leucine- and [35S]sulfate-labeled proteoglycan release of free leucine was 10 to 15 times more rapid than that of inorganic sulfate. Within approximately 3 h a steady state was approached between transport of proteoglycan to the compartment of core protein degradation and release of free leucine. No such steady state could be found with respect to the dermatan sulfate chains. In the presence of benzyloxycarbonyl-Phe-Ala-diazomethylketone or of other SH-protease inhibitors the degradation of the protein moiety of endocytosed proteoglycan was much less inhibited than the degradation of the polysaccharide chain. Benzyloxycarbonyl-Phe-Ala-diazomethylketone did not affect the degradation of dermatan sulfate chains taken up by fluid phase endocytosis and the activities of all known dermatan sulfate-degrading enzymes. Percoll gradient centrifugation indicated that also in the presence of the protease inhibitor the partially degraded proteoglycan accumulated in dense lysosomes. The isolation of intracellular dermatan sulfate peptides and molecular size determinations of endocytosed dermatan sulfate proteoglycan supported the conclusion that a critical proteolytic step is required before the dermatan sulfate chain becomes accessible to hydrolytic enzymes.
Hoppe
W
W
Institute of Physiological Chemistry and Pathobiochemistry, University of Münster, Federal Republic of Germany.
Rauch
U
U
Kresse
H
H
D000643:Ammonium Chloride / Q000494:pharmacology; D001692:Biological Transport; D002478:Cells, Cultured; D002807:Chondroitin / Q000031:analogs & derivatives*; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D003513:Cycloheximide / Q000494:pharmacology; D003871:Dermatan Sulfate / Q000378:metabolism*; D003978:Diazomethane / Q000031:analogs & derivatives / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006207:Half-Life; D006801:Humans; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D007976:Leupeptins / Q000494:pharmacology; D008247:Lysosomes / Q000378:metabolism; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism
D011508:Chondroitin Sulfate Proteoglycans; D007976:Leupeptins; D011480:Protease Inhibitors; D011509:Proteoglycans; D013431:Sulfates; C024918:benzyloxycarbonylphenylalanylalanine diazomethyl ketone; C032649:dermatan sulfate proteoglycan; D000643:Ammonium Chloride; D003871:Dermatan Sulfate; D003978:Diazomethane; D002807:Chondroitin; D003513:Cycloheximide; D007930:Leucine; C032854:leupeptin