Isolation and characterization of a peroxidase from the airway.
Sheep airway mucus can potently scavenge hydrogen peroxide, an important mediator of airway inflammation. Here, the scavenging activity was identified as a peroxidase produced by goblet cells of the airway epithelium and secreted into the airway lumen. Ovine airway peroxidase activity was purified approximately 100-fold from airway lavage fluid in two steps, using cation exchange and lectin affinity chromatography, yielding an apparently homogeneous 82-kD glycoprotein. Ovine airway peroxidase represented about 1% of the total protein in airway mucus and thus was an abundant enzyme in airway secretions. The absorbance spectrum of the purified peroxidase showed a major peak at 412 nm indicative of a hemoprotein. The ratio of A412/A280 of the purified enzyme was 0.86. The absorption spectrum of ovine airway peroxidase, its ability to oxidize halides, its sensitivity to inhibitors and its apparent molecular mass on sodium dodecyl sulfate gels showed that airway peroxidase was similar to lactoperoxidase but distinguished from myeloperoxidase, eosinophil peroxidase as well as from glutathione peroxidases. Based on these observations, ovine airway peroxidase is a newly isolated and abundant enzyme of airway mucus which may function to control reactive oxygen species in the airway and to prevent infection by catalyzing the formation of biocidal compounds.
Salathe
M
M
Division of Pulmonary and Critical Care Medicine, University of Miami School of Medicine, Florida 33136, USA.
Holderby
M
M
Forteza
R
R
Abraham
W M
WM
Wanner
A
A
Conner
G E
GE
D000818:Animals; D001992:Bronchoalveolar Lavage Fluid; D002417:Cattle; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D016166:Free Radical Scavengers; D006651:Histocytochemistry; D007700:Kinetics; D008854:Microscopy, Electron; D008970:Molecular Weight; D009092:Mucous Membrane / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure; D010544:Peroxidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012756:Sheep; D014132:Trachea / Q000201:enzymology*
D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D010544:Peroxidases