Identification and characterization of tyrosylprotein sulfotransferase from human saliva.
Tyrosylprotein sulfotransferase (TPST), the enzyme responsible for the sulfation of tyrosine residues, has been identified and characterized in submandibular salivary glands previously (William et al. Arch Biochem Biophys 338: 90-96). Tyrosylprotein sulfotransferase catalyses the sulfation of a variety of secretory and membrane proteins and is believed to be present only in the cell. In the present study, this enzyme was identified for the first time in human saliva. Analysis of human saliva and parotid saliva for the presence of tyrosylprotein sulfotransferase revealed tyrosine sulfating activity displayed by both whole saliva and parotid saliva at pH optimum of 6.8. In contrast to tyrosylprotein sulfotransferase isolated from submandibular salivary glands, salivary enzyme does not require the presence of Triton X-100, NaF and 5'AMP for maximal activity. Similar to the submandibular TPST, the enzyme from saliva also required MnCl(2) for its activity. Maximum TPST activity was observed at 20 mM MnCl(2). The enzyme from saliva was immunoprecipitated and purified by immunoaffinity column using anti-TPST antibody. Affinity purified salivary TPST showed a single band of 50-54 kDa. This study is the first report characterizing a tyrosylprotein sulfotransferase in a secretory fluid.
Kasinathan
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C
Oral Biology, NJ Dental School, University of Medicine and Dentistry of New Jersey, Newark, NJ 07103-2400, USA. kasinach@umdnj.edu
Ramaprasad
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P
Sundaram
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P
D002712:Chlorides / Q000494:pharmacology; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007700:Kinetics; D017895:Manganese Compounds / Q000494:pharmacology; D010306:Parotid Gland / Q000201:enzymology / Q000378:metabolism; D012016:Reference Values; D012463:Saliva / Q000201:enzymology* / Q000378:metabolism; D015238:Sulfotransferases / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D002712:Chlorides; D017895:Manganese Compounds; D014443:Tyrosine; D015238:Sulfotransferases; C039079:protein-tyrosine sulfotransferase; C025340:manganese chloride