Abstract
Human cytosolic sulfotransferase SULT1E1 catalyzes the sulfation of endogenous estrogens as well as xenobiotic estrogen-like chemicals. This reaction increases the water solubility of the molecule, which may affect its cellular distribution and biological activity. This could alter estrogen signaling to the estrogen receptor in human estrogen receptor-positive cells. The current work characterized the cellular distribution of SULT1E1 in the human embryonic kidney 293 (HEK293) cell line using green fluorescent protein (GFP) tagging and immunochemistry methods. The GFP-tagged recombinant SULT1E1 protein was expressed and localized in the cytoplasm of HEK293 cells. By using a commercial anti-SULT1E1 peptide antibody, a 35.7-kDa protein was detected in HEK293 cells via Western blot. The molecular mass of the protein detected suggested that it may be related to native SULT1E1 protein. However, reverse transcription-polymerase chain reaction (RT-PCR) with gene-specific primers could not confirm the presence of the SULT1E1 transcript in the total RNA sample of HEK293 cells. The discrepancy between protein and transcript data could be due to the instability of SULT1E1 mRNA or the specificity of the anti-SULT1E1 antibody used. In the present work, RT-PCR analysis with gene-specific primers also identified a transcript fragment of human estrogen-related receptor γ. Future studies on the functional relationship between estrogen-related receptors and sulfotransferases are expected to provide additional insights into the physiological and toxicological roles of human estrogen sulfotransferases.
Footnotes
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This work was supported by North Dakota Biomedical Research Infrastructure Network (ND BRIN), North Dakota State University, Fargo, ND.
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Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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doi:10.1124/dmd.106.011247.
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ABBREVIATIONS: SULT1E1, human estrogen sulfotransferase 1E1; ER-α, estrogen receptor α; ER-β, estrogen receptor β; BSA, bovine serum albumin; ERR-γ, estrogen-related receptor γ; GFP, green fluorescent protein; HEK293, human embryonic kidney 293; PBS, phosphate-buffered saline; RT-PCR, reverse transcription-polymerase chain reaction.
- Received June 5, 2006.
- Accepted October 5, 2006.
- The American Society for Pharmacology and Experimental Therapeutics
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