@article {886,
	title = {Identification and purification of sulfotransferases for 20-hydroxysteroid from the larval fat body of a fleshfly, Sarcophaga peregrina},
	volume = {67},
	year = {Submitted},
	note = {08-10-200906-02-2012NormalKyoritsu College of Pharmacy, Minato-ku, Tokyo 105-8512, Japan.enghttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=12951514},
	pages = {1780 - 1785},
	abstract = {Sulfotransferase (ST) activity for 20-hydroxyecdysone (20E) was identified in a larval fat body lysate of the fleshfly, Sarcophaga peregrina, but not in the hemolymph. The activity was highly sensitive to 2,6-dichloro-4-nitrophenol (DCNP) (IC50=0.61 microM), a specific inhibitor of phenol ST (P-ST), but insensitive to triethylamine, a hydroxysteroid ST inhibitor. These results suggest that 20E-specific ST enzymes belong to the P-ST family, despite the fact that 20E is a hydroxysteroid. In addition to 20E ST activity, a relatively high level of 2-naphthol ST activity was detected in the fat body lysate. The ST activity for both substrates transiently decreased to the 50\% of maximal levels, 6 hrs after induction of pupation. The ST enzymes were separated on a DEAE-cellulose column. The 20E-ST enzymes were eluted around 50 mM KCl as two separate peaks of close proximity and the P-ST was eluted at 0.1 M KCl. The 20E ST enzymes were further purified using 3{\textquoteright}-phosphoadenosine 5{\textquoteright}-phosphate (PAP)-agarose affinity column chromatography. Both of the eluted active fractions demonstrated 43-kDa proteins on SDS-polyacrylamide gel. Photoaffinity labeling with [35S]-3{\textquoteright}-phosphoadenosine 5{\textquoteright}-phosphosulfate (PAPS) showed 43-kDa bands in the fat body lysate, as well as in the purified fractions. These results suggest that the 43-kDa proteins catalyze 20E sulfation within the fat body of S. peregrina.},
	keywords = {Adenosine Diphosphate/pharmacology, Animals, Chromatography, DEAE-Cellulose, derivatives/metabolism, development, Diptera/*enzymology/growth \&amp, Ecdysterone/antagonists \&amp, Enzyme Inhibitors/chemistry/pharmacology, Fat Body/chemistry/*enzymology, Hemolymph/metabolism, inhibitors/*chemistry/isolation \&amp, inhibitors/isolation \&amp, Larva/chemistry/enzymology, Naphthols/isolation \&amp, Nitrophenols/pharmacology, Phosphoadenosine Phosphosulfate/analogs \&amp, Photoaffinity Labels/chemistry/metabolism, Pupa/metabolism, purification/*metabolism, purification/metabolism, Sulfates/metabolism, Sulfotransferases/antagonists \&amp, Sulfur Radioisotopes},
	author = {Matsumoto, E. and Matsui, M. and Tamura, H. O.}
}
