Cloning, expression, and characterization of cytosolic sulfotransferase isozymes from Drosophila melanogaster.
We have identified four cytosolic sulfotransferase (SULT) homologs in the genome database of Drosophila melanogaster, and have designated these genes dmST1-4. Each of these four isozymes was subsequently classified into a different and novel gene family, as determined by the low amino acid sequence homology (less than 40%) between them, and also toward their vertebrate homologs. The transcripts for these four SULT homologs were detectable at all developmental stages in D. melanogaster. In addition, three of these isozymes, the exception being dmST2, were successfully expressed and purified in Escherichia coli. These recombinant dmST1, 3, and 4 products showed high sulfating activity toward phenolic compounds such as vanillin and 4-nitrophenol, but showed no activity toward typical endogenous substrates such as tyramine and serotonin. DmST4 and dmST3 showed the lowest and highest K(m) values for 3'-phosphoadenosine-5'-phosphosulfate (PAPS) respectively. DmST4 also showed low but not negligible activity toward 20-hydroxyecdysone.
Hattori
Kenji
K
Kyoritsu University of Pharmacy, Tokyo 105-8512, Japan.
Motohashi
Noriko
N
Kobayashi
Izumi
I
Tohya
Takeshi
T
Oikawa
Mai
M
Tamura
Hiro-omi
HO
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology*; D017931:DNA Primers; D004331:Drosophila melanogaster; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D015238:Sulfotransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D015238:Sulfotransferases