Significance of selenium-labeled proteins for selenium's chemopreventive functions.
A 58 kDa selenium-labeled protein purified from mouse mammary epithelial cells (MMEC) was used to examine whether selenium modulates protein synthesis or is just a marker for cellular selenium status. The protein was isolated using Sephadex G150 gel filtration and DEAE-Sephadex A50 ion-exchange chromatography. It was further analysed using 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and was found as a single spot with a pI of 4.6. The immunoreactivity with anti-58 kDa antiserum and the 75Se signal co-localized on a single 58 kDa protein band on both 1D- and 2D-PAGE. Partial amino acid analysis of the peptide showed homology with the thiol protein disulfide oxidoreductase (TPDO). Varying the selenium concentration in culture medium did not affect the protein content or the immunoreactivity of the 58 kDa protein. Additionally, selenium did not seem to regulate the activity of TPDO in TM6 cells. The glutathione peroxidase activity of TM6 cells, taken as the internal positive control, was enhanced with the increase in selenium concentration in the medium. The results suggest that selenium is attached to the 58 kDa protein, but does not regulate either its protein synthesis or its functional activity. We conclude that selenium labeling of the 58 kDa protein reflects the cellular selenium status but probably is not involved in its chemopreventive ability.
Sinha
R
R
Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030.
Bansal
M P
MP
Ganther
H
H
Medina
D
D
D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005979:Glutathione Peroxidase / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011490:Protein Disulfide Reductase (Glutathione) / Q000737:chemistry / Q000302:isolation & purification*; D012643:Selenium / Q000378:metabolism / Q000494:pharmacology*; D012644:Selenium Radioisotopes / Q000378:metabolism
D012644:Selenium Radioisotopes; D005979:Glutathione Peroxidase; D011490:Protein Disulfide Reductase (Glutathione); D012643:Selenium