Identification of Two Human Brain Aryl Sulfotransferase cDNAs

Molecular cloning, expression, localisation and functional characterisation of a rabbit SULT1C2 sulfotransferase

“…Control assays using COS cells transfected with vector only were undertaken to confirm a lack of endogenous activity. A range of potential substrates that have previously been shown to be sulfonated by humSULT1A1 [21], humSULT1A3 [22], humSULT2A1 [23] and humSULT1E [24] were tested. In total, 16 different compounds were screened using a range of concentrations (both in micromolar and millimolar ranges) as potential substrates; 4-acetaminophenol, allopurinol, 7-OH-2-acetylaminofluorine (AAF), bromophenol blue, 4-chloro-1-napthol, chlorozoxazone, dexamethasone, DHEA, dopamine, estrone, 4-hydroxytamoxifen, indole-3-carbinol, minoxidil, p-nitrophenol, poly-EAY (Glu, Ala, Tyr, 6:3:1) and thyroxine.…”

Bacterial expression of two human aryl sulfotransferases

“…Bacteria were cultured for 24 h at 30°C with 1 mM IPTG and subcellular fractions were prepared as described previously [8]. Lanes 1 and 2, 50 μg of total protein from COS cells transfected with pCMV5/HAST1 and pCMV5/HAST3, respectively [2,3]. The functional characteristics of E. coli-expressed HAST1 and HAST3 proteins were determined towards their model substrates p-nitrophenol and dopamine, respectively, using a modified procedure of Foldes and Meek [10,11].…”

“…These include five aryl sulfotransferases, a hydroxysteroid sulfotransferase and an estrogen sulfotransferase [1]. In our laboratory, five human aryl sulfotransferase (HAST) cDNAs have been cloned (HAST1, HAST2, HAST3, HAST4 and HAST4v) [2,3,4]. HAST1 and HAST2 are thermostable forms of aryl sulfotransferases and sulfonate small phenolic compounds such as p-nitrophenol.…”

“…HAST1 and HAST2 are thermostable forms of aryl sulfotransferases and sulfonate small phenolic compounds such as p-nitrophenol. The cDNAs which encode these proteins are identical in their coding sequence but differ in the 5′ untranslated regions [2,3]. HAST3, a thermolabile form of aryl sulfotransferase is 93% identical in its coding sequence to HAST1 and HAST2.…”

“…HAST3, a thermolabile form of aryl sulfotransferase is 93% identical in its coding sequence to HAST1 and HAST2. The monoamine dopamine is the preferred substrate of HAST3 [3,5]. Recently we have cloned two cDNAs which we have termed HAST4 and HAST4v [4].…”