Inhibition of leucocytic lysosomal enzymes by glycosaminoglycans in vitro.
1. A lysosomal fraction was separated by density-gradient centrifugation from a highly purified human polymorphonuclear leucocyte suspension. 2. Some 23 different lysosomal enzymes were assayed for activity in the presence of various concentrations of glycosaminoglycans. 3. The 21 acid hydrolases assayed were strongly inhibited to different degrees by low (0-12 mmol/l) concentrations of glycosaminoglycans in a pH-dependent manner. Thus inhibitions were stronger below pH4.5, with activity returning to control values at about pH5.0. 4. On a molar basis, the inhibitory activity for the several glycosaminoglycans studied was: heparin greater than chondroitin sulphate greater than hyaluronic acid. 5. Once the glycosaminoglycan-acid hydrolase complex was formed, it was partially dissociated by slight elevations in the pH of the incubation medium, by increasing the ionic strength of the incubation medium, or by adding several cationic proteins (e.g. histone, protamine). 6. As leucocytic lysosomes contain large amounts of chondroitin sulphate, and have a strongly acid intragranular pH, we suggest that glycosaminoglycans may modify lysosomal function through the formation of complexes with lysosomal enzymes, by inhibiting the digestive activity of the acid hydrolases when the intralysosomal pH is below their pI.
D000135:Acid Phosphatase / Q000037:antagonists & inhibitors; D001192:Arylsulfatases / Q000037:antagonists & inhibitors; D002809:Chondroitin Sulfates / Q000494:pharmacology; D006025:Glycosaminoglycans / Q000494:pharmacology*; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors; D006493:Heparin / Q000494:pharmacology; D006657:Histones / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007962:Leukocytes / Q000187:drug effects / Q000201:enzymology*; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology*; D010544:Peroxidases / Q000037:antagonists & inhibitors; D010740:Phospholipases / Q000037:antagonists & inhibitors; D011402:Pronase / Q000032:analysis; D011479:Protamines / Q000494:pharmacology; D011480:Protease Inhibitors; D012260:Ribonucleases / Q000037:antagonists & inhibitors
D006025:Glycosaminoglycans; D006657:Histones; D011479:Protamines; D011480:Protease Inhibitors; D006493:Heparin; D002809:Chondroitin Sulfates; D010544:Peroxidases; D010740:Phospholipases; D012260:Ribonucleases; D000135:Acid Phosphatase; D001192:Arylsulfatases; D006026:Glycoside Hydrolases; D011402:Pronase