Rapid sulfation of 3,3′,5′-triiodothyronine in native Xenopus laevis oocytes

Sulfation is an important metabolic pathway facilitating the degradation of thyroid hormone by the type I iodothyronine deiodinase. Different human and rat tissues contain cytoplasmic sulfotransferases that show a substrate preference for 3,3'-diiodothyronine (3,3'-T-2) > T-3 > rT(3) > T-4. During investigation of the expression of plasma membrane transporters for thyroid hormone by injection of rat liver RNA in Xenopus laevis oocytes, we found uptake and metabolism of iodothyronines by native oocytes. Groups of 10 oocytes were incubated for 20 h at 18 C in 0.1 mi medium containing 500,000 cpm (1-5 nM) [I-125]T-4, [I-125]T-3, [I-125]rT(3), or [I-125]3,3'-T-2. In addition, cytosol prepared from oocytes was tested for iodothyronine sulfotransferase activity by incubation of 1 mg cytosolic protein/ml for 30 min at 21 C with 1 mu M [I-125]T-4, [I-125]T-3, [I-125]rT(3), or [I-125]3,3'-T-2 and 50 mu M 3'-phosphoadenosine-5'-phosphosulfate. Incubation media, oocyte extracts, and assay mixtures were analyzed by Sephadex LH-20 chromatography for production of conjugates and iodide. After 20-h incubation, the percentage of added radioactivity present as conjugates in the media and oocytes amounted to 0.9 +/- 0.2 and 1.0 +/- 0.1 for T-4, less than 0.1 and less than 0.1 for T-3, 32.5 +/- 0.4 and 29.3 +/- 0.2 for rT(3), and 3.8 +/- 0.3 and 2.3 +/- 0.2 for 3,3'-T-2, respectively (mean +/- SEM; n = 3). The conjugate produced from rT(3) was identified as rT, sulfate, as it was hydrolyzed by acid treatment. After injection of oocytes with copy RNA coding for rat type I iodothyronine deiodinase, we found an increase in iodide production from rT(3) from 2.3% (water-injected oocytes) to 46.2% accompanied by a reciprocal decrease in rT(3) sulfate accumulation from 53.7% to 7.1%. After 30-min incubation with cytosol and 3'-phosphoadenosine-5'-phosphosulfate, sulfate formation amounted to 1.8% for T-4, less than 0.1% for T-3, 77.9% for rT(3), and 2.9% for 3,3'-T-2. These results show that rT(3) is rapidly metabolized in native oocytes by sulfation. The substrate preference of the sulfotransferase activity in oocytes is rT(3) much greater than 3,3'-T-2 > T-4 > T-3. The physiological significance of the high activity for rT(3) sulfation in X. laevis oocytes remains to be established.