Inhibition by heparin of the oxidation of lysine in collagen by lysyl oxidase.
The generation of covalent cross-linkages in collagen is initiated by the deamination by lysyl oxidase of specific lysine residues in this connective tissue protein. Since lysyl oxidase activity is influenced by ionic ligands bound to its protein substrates, the effect of heparin, an anionic glycosaminoglycan known to bind to collagen, was explored by using collagen and elastin substrates and highly purified lysyl oxidase. Concentrations of heparin up to 1 mg mL-1 had little effect on the enzymatic rate of oxidation if it was added prior to the addition of enzyme to a preformed fibrillar collagen substrate or to an insoluble elastin substrate. However, collagen oxidation was inhibited by 85% if this glycosaminoglycan was present at 0.4 mg mL-1 during collagen fibril formation before addition of the enzyme. Similarly, the rate and extent of collagen fibrillogenesis in the absence of lysyl oxidase were each markedly inhibited in the presence of 0.4 mg mL-1 heparin. Heparin also inhibited the extent of tight binding of lysyl oxidase to preformed fibrils by about 40% under conditions where enzyme activity against preformed fibrils was hardly affected. These results suggest that heparin may modulate the oxidation and thus the insolubilization of extracellular collagen fibers, possibly under conditions where elastin fiber synthesis is not affected, and that the tight binding of lysyl oxidase to collagen is not completely related to the expression of enzyme activity toward this substrate. These results also have mechanistic implications for the retarding effect of heparin on postoperative wound healing.
D000594:Amino Acid Oxidoreductases / Q000037:antagonists & inhibitors*; D000818:Animals; D001011:Aorta / Q000201:enzymology; D002417:Cattle; D003094:Collagen / Q000378:metabolism*; D003432:Cross-Linking Reagents; D006493:Heparin / Q000494:pharmacology*; D007700:Kinetics; D008239:Lysine / Q000378:metabolism*; D010084:Oxidation-Reduction; D011485:Protein Binding; D008249:Protein-Lysine 6-Oxidase / Q000037:antagonists & inhibitors*
D003432:Cross-Linking Reagents; D006493:Heparin; D003094:Collagen; D000594:Amino Acid Oxidoreductases; D008249:Protein-Lysine 6-Oxidase; D008239:Lysine