Purification from rat liver of an enzyme that catalyses the sulphurylation of phenols

Abstract

1. An enzyme (EC 2.8.2.1) that catalyses the transfer of sulphate from adenosine 3'-phosphate 5'-sulphatophosphate to phenols was purified approx. 2000-fold from male rat livers. 2. The purified preparation did not catalyse the sulphurylation of dehydroepiandrosterone, butan-1-ol, l-tyrosine methyl ester, 1-naphthylamine or serotonin. 3. At pH8.0 and 37 degrees C the K(m) values of the enzyme for p-nitrophenol and adenosine 3'-phosphate 5'-sulphatophosphate are 51 and 14mum respectively. The K(m) value for either substrate is independent of the concentration of the other. 4. The sulphurylation of phenol is inhibited by thiol compounds and glutathione at a concentration of 3mm caused an approx. 50% decrease in enzyme activity. 5. The K(m) of the enzyme for adenosine 3'-phosphate 5'-sulphatophosphate is unaffected by the presence of added glutathione but at a concentration of 5mm-glutathione the K(m) of the enzyme for its phenolic substrate is decreased.

Phenol sulphotransferase and uridine diphosphate glucuronyltransferase from rat liver in vivo and in vitro. 2,6-Dichloro-4-nitrophenol as selective inhibitor of sulphation

“…phenol or N-hydroxyphenacetin. As sulphotransferase preparation a partly purified rat liver phenol sulphotransferase was used, purified by the method of McEvoy & Carroll (1971) as far as the (NH4)2SO4 precipitation and subsequent dialysis.…”

Enzymic sulphation of p-nitrophenol and steroids by larval gut tissues of the southern armyworm (Prodenia eridania Cramer)

“…Although sulphate conjugates of phenol were first isolated and identified in mammals im the latter part ofthe last century, the mechanism ofthis reaction was not fully established until the discovery ofthe enzymic activation of sulphate and the isolation and identification of active sulphate, 3'-phosphoadenylyl sulphate (adenosine 3'-phosphate 5'-sulphatophosphate) some 15-20 years ago (Bernstein & McGilvery, 1952;DeMeio et al, 1953;Segal, 1955;DeMeio et al, 1955;Hilz & Lipmann, 1955;Robbins & Lipmann, 1956a,b, 1957. Studies on sulphate transfer from 3'phosphoadenylyl sulphate to various acceptors including mammalian hormonal steroids and L-tyrosine methyl ester have been actively pursued in the last few years. In mammals much of the recent emphasis on sulphoconjugation has been centred around the purification and characterization of the individual enzymes within the complex-forming family of sulphotransferases (Nose & Lipmann, 1958;Banerjee & Roy, 1966, 1968Hidaka et al, 1969;Mattock & Jones, 1970;McEvoy & Carroll, 1971). It now appears that in mammalian tissues there are at least four different sulphotransferases (Dodgson & Rose, 1970;Roy, 1970), which according to acceptor specificity are designated as phenol sulphotransferase (3'-phosphoadenylyl sulphate-phenol sulphotransferase, EC 2.8.2.1), 3fl-hydroxy steroid sulphotransferase (3'phosphoadenylyl sulphate-3p-hydroxy steroid sulphotransferase, EC 2.8.2.2), oestrone sulphotransferase (3'-phosphoadenylyl sulphate-oestrone sulphotransferase, EC 2.8.2.4) and L-tyrosine methyl ester sulphotransferase.…”

“…Of the enzymes that have been partially purified, phenol sulphotransferase is reported to have been purified more than 2000-fold Vol. 130 from male rat liver (McEvoy & Carroll, 1971). This purified enzyme did not catalyse the sulphation of dehydroepiandrosterone, butan-1 -ol, L-ttyrosine methyl ester, 1-naphthylamine or 5-hydroxytryptamine (serotonin).…”