Abstract
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 μM for bronchial, 31.3 μM for parenchymal). Anion exchange chromatography indicated two PST isoforms being expressed in different ratios. Immunoblot analysis with mouse antibovine 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.
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References
Adams JB, McDonald D (1981) Enzymic synthesis of steroid sulphates. XIV. Properties of human adrenal steroid alcohol sulphotransferase. Biochim Biophys Acta 664:460–468
Arand M, Robertson LW, Oesch F (1987) A fluorometric assay for quantitating phenol sulfotransferase activities in homogenates of cells and tissues. Anal Biochem 163:546–551
Autrup H, Wefald F, Jeffrey AM, Tate H, Schwartz RD, Trump BF, Harris CC (1980) Metabolism of benzo(a)pyrene by cultured tracheobronchial tissues from mice, rats, hamsters, bovines and humans. Int J Cancer 25:293–300
Baranczyk-Kuzma A, Szymczyk T (1986) Lung phenol sulfotransferases: thermal stability of human and bovine enzymes. Biochem Pharmacol 35:995–999
Baranczyk-Kuzma A, Szymczyk T (1987) Extrahepatic sulfation of phenols: bovine lung and intestinal phenol sulfotransferases. Biochem Pharmacol 36:3141–3146
Beckmann JD (1991) Continuous fluorometric assay of phenol sulfotransferase. Anal Biochem 197:408–411
Beckmann JD, Takizawa H, Romberger D, Illig M, Claassen L, Rickard K, Rennard SI (1992) Serum free culture of fractionated bovine bronchial epithelial cells. In Vitro Cell Dev Biol 28A:39–46
Culling CEA, Allison RT, Barr WT (1985) Cellular pathology technique. Butterworth, London
Jeffery PK (1983) Morphologic features of airway surface epithelial cells and glands. Am Rev Respir Dis 128:S14-S20
Johnson GD, Nogueira Araujo GM (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods 43:349–350
Kane RE, Chen LF, Thaler MM (1984) Regulation of bile salt sulfotransferase isoenzymes by gonadal hormones. Hepatology 4:1195–1199
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685
Lin YN, Horowitz MI (1980) Enzymatic sulfation of cholesterol by rat gastric mucosa. Steroids 36:697–708
Markwell MA, Haas SM, Bieber LL, Tolbert NE (1978) A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87:206–210
Mulder GJ, Scholtens E (1977) Phenol sulphotransferase and uridine diphosphate glucuronyltransferase from rat liver in vivo and in vitro. Biochem J 165:553–559
Nemoto N (1981) Glutathione, glucuronide, and sulfate transferase in polycyclic aromatic hydrocarbon metabolism. Polycyclic Hydrocarbons Cancer 3:213–258
Overby L, Nishio SJ, Lawton MP, Plopper CG, Philpot RM (1992) Cellular localization of flavin-containing monooxygenase in rabbit lung. Exp Lung Res 18:131–144
Ramaswamy SG, Jakoby WB (1987) Sulfotransferase assays. Methods Enzymol 143:201–207
Rearick JI, Albro PW, Jetten AM (1987) Increase in cholesterol sulfotransferase activity during in vitro squamous differentiation of rabbit tracheal epithelial cells and its inhibition by retinoic acid. J Biol Chem 26:13069–13074
Rens-Domiano S, Roth JA (1989) Characterization of tyrosylprotein sulfotransferase from rat liver and other tissues. J Biol Chem 264:899–905
Roy AB (1960) The enzymic synthesis of aryl sulphamates. Biochemistry 74:49–56
Roy AB (1981) Sulfotransferases. In: Mulder GJ (ed) Sulfation of drugs and related compounds, pt 2. CRC Press, Boca Raton, Fla, pp 83–130
Sekura RD, Jakoby WB (1979) Phenol sulfotransferases. J Biol Chem 254:5658–5663
Sekura RD, Marcus CJ, Lyon ES, Jakoby WB (1979) Assay of sulfotransferases. Anal Biochem 95:82–86
Sims P, Grover PL (1974) Epoxides in polycyclic aromatic hydrocarbon metabolism and carcinogenesis. Adv Cancer Res 20:165–274
Spina D, Rigby PJ, Paterson JW, Goldie RG (1989) Autoradiographic localization of beta-adrenoceptors in asthmatic human lung. Am Rev Respir Dis 140:1410–1415
Sundaram RS, Szumlanski C, Otterness D, Van Loon JA, Weinshilboum RM (1989) Human intestinal phenol sulfotransferase: assay conditions, activity levels and partial purification of the thermolabile form. Drug Metab Dispos 17:255–264
Surh Y-J, Blomquist JC, Liem A, Miller JA (1990) Metabolic activation of 9-hydroxymethyl-10-methylanthracene and 1-hydroxymethylpyrene to electrophilic, mutagenic and tumorigenic sulfuric acid esters by rat hepatic sulfotransferase activity. Carcinogenesis 11:1451–1460
Takizawa H, Romberger D, Beckmann J, Rickard K, Matsuda T, Shoji S, Claassen L, Linder J, Rennard SI (1990) Separation of subpopulations of bovine bronchial epithelial cells by density centrifugation. Am Rev Respir Cell Mol Biol 3:553–562
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Vaught JB, McGarvey PB, Lee M-S, Garner CD, Wang CY, Linsmaier-Bednar EM, King CM (1981) Activation of N-hydroxyphenacetin to mutagenic and nucleic acid-binding metabolites by acyltransfer, deacylation, and sulfate conjugation. Cancer Res 41:3424–3429
Walle T, Walle UK (1992) Stereoselective sulfate conjugation of 4-hydroxypropanolol and terbutaline by the human liver phenolsulfotransferases. Drug Metab Dispos 20:333–336
Watabe T, Fujieda T, Hiratsuka A, Ishizuka T, Hakamata Y, Ogura K (1985) The carcinogen, 7-hydroxymethyl-12-methylbenz(a)anthracene, is activated and covalently binds to DNA via a sulphate ester. Biochem Pharmacol 34:3002–3005
Wiebel FJ, Kiefer F, Krupski G, Schuller HM (1986) Expression of glutathione S-transferase and phenol sulfotransferase, but not of UDP-glucuronosyltransferase, in the human lung tumor cell lines, NCI-H322 and NCI-H358. Biochem Pharmacol 35:1337–1343
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Beckmann, J.D., Spurzem, J.R. & Rennard, S.I. Phenol sulfotransferase expression in the airways: enzymological and immunohistochemical demonstration. Cell Tissue Res 274, 475–485 (1993). https://doi.org/10.1007/BF00314544
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DOI: https://doi.org/10.1007/BF00314544