Oxidative degradation of rat mast-cell heparin proteoglycan.
The susceptibility of rat mast-cell heparin to oxidative degradation was examined. Heparin as a component of intact mast-cell granules (MCG) was degraded following ingestion by normal human neutrophils. In contrast, neutrophils from patients with chronic granulomatous disease (CGD), which do not respond to stimulation with respiratory-burst activity, exhibited a greatly diminished ability to degrade phagocytosed MCG heparin. MCG-associated heparin also was cleaved by H2O2 plus Fe2+ (Fenton's reagent). Isolated heparin proteoglycan (average Mr approx. 750,000) was rapidly cleaved to smaller molecules similar in size to commercial pig heparin upon exposure to Fenton's reagent. This cleavage was inhibited by catalase and by the hydroxyl-radical (OH.)-scavenger mannitol, but not by superoxide dismutase (SOD). The cleavage products retained approx. 26% of the anticoagulant activity of the native molecule. The heparin proteoglycan was also cleaved by acetaldehyde/xanthine oxidase/FeSO4, a system that generates superoxide (O2.-), H2O2 and OH.. Whereas the cleavage at relatively high iron ion concentrations was inhibited by catalase and mannitol but not by SOD, at lower iron ion concentrations the cleavage was inhibited by catalase, mannitol and SOD. These findings suggest the involvement of OH., which at high Fe2+ concentrations is generated by Fenton's reagent (H2O2 plus Fe2+), and at low iron ion concentrations is generated by the iron-ion-catalysed interaction between O2.- and H2O2 (Haber-Weiss reaction). These studies suggest that oxygen radicals generated by activated phagocytes may contribute to the degradation in vivo of both solubilized and granule-associated proteoglycan heparin.
Metcalfe
D D
DD
Mast Cell Physiology Section, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Thompson
H L
HL
Klebanoff
S J
SJ
Henderson
W R
WR
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006105:Granulomatous Disease, Chronic / Q000097:blood; D006493:Heparin / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008297:Male; D008407:Mast Cells / Q000378:metabolism*; D009504:Neutrophils / Q000378:metabolism*; D010084:Oxidation-Reduction; D011509:Proteoglycans / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values; D013481:Superoxides / Q000378:metabolism
D011509:Proteoglycans; C036090:heparin proteoglycan; D013481:Superoxides; D006493:Heparin