Molecular cloning and characterization of rat ST1B1 and human ST1B2 cDNAs, encoding thyroid hormone sulfotransferases.
Human and rat cDNAs encoding thyroid hormone sulfotransferases have been isolated from their liver cDNA libraries. The isolated sulfotransferases, termed rat ST1B1 and human ST1B2, share 77 and 74% homologies at nucleotide and deduced amino acid levels. These forms showed less than 36 and 56% homologies to hydroxysteroid and aryl sulfotransferases, indicating that they constitute a new gene subfamily of aryl sulfotransferase. Expression of ST1B1 and ST1B2 in COS-1 cells resulted in the appearance of 33.0 and 32.5 kDa proteins, respectively, whose mobilities were identical with proteins detected in rat and human livers in Western blots using antibodies raised against ST1B1 and ST1B2 produced in Escherichia coli. The recombinant forms catalyzed sulfation of p-nitrophenol, 3,3',5-triiodothyronine (T3) and dopamine, but not of beta-estradiol and dehydroepiandrosterone. ST1B1 and ST1B2 showed higher affinities for formation of T3 sulfate (apparent Km 40.2 and 63.5 microM, respectively) than did thermostable phenol sulfotransferase ST1A3 (apparent Km 413 microM) or thermolabile phenol sulfotransferase ST1A5 (apparent Km 180 microM). These data indicate that the newly characterized sulfotransferases constitute a distinct ST1 subfamily of enzymes catalyzing the sulfation of T3 as a typical endogenous substrate in rats and humans.
Fujita
K
K
Division of Drug Metabolism and Molecular Toxicology, Faculty of Pharmaceutical Sciences, Tohoku University, Sendai.
Nagata
K
K
Ozawa
S
S
Sasano
H
H
Yamazoe
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D002384:Catalysis; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017386:Sequence Homology, Amino Acid; D015238:Sulfotransferases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D014284:Triiodothyronine / Q000378:metabolism*
D018076:DNA, Complementary; D012333:RNA, Messenger; D014284:Triiodothyronine; D015238:Sulfotransferases; C109377:iodothyronine sulfotransferase