Phenol sulfotransferase expression in the airways: enzymological and immunohistochemical demonstration.
Phenol (aryl) sulfotransferases (PSTs) provide a conjugative pathway that detoxifies hydroxylated aromatic xenobiotics by esterification with sulfate. Both human and bovine airways have been reported to use this pathway, and in this investigation the bovine system is examined. PST activity in tracheal through fourth generation bronchial mucosal cytosols was 0.1-0.35 nmol/mg protein/min. Activity was generally greater in more distal bronchi and in parenchymal extracts, which contained 0.6-3 nmol/mg/min PST activity. Comparison of the PST activities of bronchial and parenchymal cytosols indicated similar pH activity profiles, although steady state kinetic measurements revealed different Km values for the acceptor substrate 2-naphthol (13.7 microM for bronchial, 31.3 microM for parenchymal). Anion exchange chromatography indicated two PST isoforms being expressed in different ratios. Immunoblot analysis with mouse anti-bovine PST revealed a closely spaced doublet at 32 kDa in both bronchial mucosal and parenchymal cytosolic extracts; however, this doublet was unequally stained in parenchymal extracts. Immunohistochemical analyses revealed faint positive staining of the tracheobronchial epithelium. Greatest immunostaining was observed in the nonciliated secretory epithelial cells of the bronchioles, whereas surrounding smooth muscle, endothelial cells, and alveoli were immunonegative. These results are consistent with the known locations of other detoxification enzymes within the airways.
Beckmann
J D
JD
Department of Internal Medicine Pulmonary and Critical Care, University of Nebraska Medical Center, Omaha 68198.
Spurzem
J R
JR
Rennard
S I
SI
D000818:Animals; D015239:Arylsulfotransferase / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D015153:Blotting, Western; D001980:Bronchi / Q000166:cytology / Q000201:enzymology*; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006863:Hydrogen-Ion Concentration; D013463:Sulfuric Acid Esters / Q000032:analysis; D014132:Trachea / Q000166:cytology / Q000201:enzymology*
D013463:Sulfuric Acid Esters; C029092:naphthyl sulfate; D015239:Arylsulfotransferase