Selenoproteins are expressed in fetal human osteoblast-like cells.

Selenoproteins are involved in mechanisms of cell differentiation and defense. We investigated the expression of glutathione peroxidases, as well as other selenoproteins, in fetal human osteoblasts (hFOB-cells). Using 75-selenium metabolic labelling of viable hFOB-cells, we identified several selenoproteins in cell lysates of about 45-80 kDa and in the migration range of 14 kDa to 24 kDa. Cells expressed low mRNA levels of both cellular glutathione peroxidase and plasma glutathione peroxidase mRNA as analysed by Southern analysis of RT-PCR products. Basal cellular glutathione peroxidase enzyme activity in hFOB-cells (19.7 nmol NADPH oxidised per min and microg protein) was further increased 2.5-fold by the addition of 100 nM sodium selenite to the culture medium for 3 days. Furthermore, expression of selenoprotein P mRNA was demonstrated by RT-PCR. hFOB-cells did not show activities of the selenoproteins type I or type II 5'-deiodinase. In summary, we identified cellular glutathione peroxidase, plasma glutathione peroxidase and selenoprotein P among of a panel of several 75-selenium labelled proteins in human fetal osteoblasts. The expression of selenoproteins like glutathione peroxidases in hFOB-cells represents a new system of osteoblast antioxidative defense that may be relevant for the protection against hydrogen peroxide produced by osteoclasts during bone remodelling.

[1]  Tatjana C. Jakobs,et al.  Cloning and Characterization of the Human Selenoprotein P Promoter , 1997, The Journal of Biological Chemistry.

[2]  K. Saijoh,et al.  Analysis of bovine selenoprotein P-like protein gene and availability of metal responsive element (MRE) located in its promoter. , 1997, Gene.

[3]  R. Brigelius-Flohé,et al.  Determinants of PHGPx expression in a cultured endothelial cell line. , 1997, Biomedical and environmental sciences : BES.

[4]  R. Schirmer,et al.  The mechanism of thioredoxin reductase from human placenta is similar to the mechanisms of lipoamide dehydrogenase and glutathione reductase and is distinct from the mechanism of thioredoxin reductase from Escherichia coli. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[5]  M. Brandi,et al.  A Human Homolog of the 150 kD Avian Osteoclast Membrane Antigen Related to Superoxide Dismutase and Essential for Bone Resorption is Induced by Developmental Agents and Opposed by Estrogen in FLG 29.1 Cells , 1997, Calcified Tissue International.

[6]  C. Marcocci,et al.  Skeletal Integrity in Men Chronically Treated with Suppressive Doses of L‐Thyroxine , 1997, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[7]  L. Mosekilde,et al.  Is Skeletal Responsiveness to Thyroid Hormone Altered in Primary Osteoporosis or Following Estrogen Replacement Therapy? , 1997, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[8]  N. Copeland,et al.  Identification of a novel selD homolog from eukaryotes, bacteria, and archaea: is there an autoregulatory mechanism in selenocysteine metabolism? , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J. Compston,et al.  The expression of thyroid hormone receptors in human bone. , 1996, Bone.

[10]  J H Fraser,et al.  Hydrogen peroxide, but not superoxide, stimulates bone resorption in mouse calvariae. , 1996, Bone.

[11]  R. Rodriguiz,et al.  Osteoclastic superoxide production and bone resorption: Stimulation and inhibition by modulators of NADPH oxidase , 1996, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[12]  E. Westhof,et al.  A novel RNA structural motif in the selenocysteine insertion element of eukaryotic selenoprotein mRNAs. , 1996, RNA.

[13]  Q. Sun,et al.  Expression of human phospholipid hydroperoxide glutathione peroxidase gene for protection of host cells from lipid hydroperoxide-mediated injury. , 1996, Biochemical and biophysical research communications.

[14]  T. Stadtman,et al.  A new selenoprotein from human lung adenocarcinoma cells: purification, properties, and thioredoxin reductase activity. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[15]  G. D. Markham,et al.  Osteoclast radical interactions: NADPH causes pulsatile release of NO and stimulates superoxide production. , 1995, Endocrinology.

[16]  G. Powis,et al.  Cloning and sequencing of a human thioredoxin reductase , 1995, FEBS letters.

[17]  P. Newburger,et al.  Structure and function of the selenium translation element in the 3'-untranslated region of human cellular glutathione peroxidase mRNA. , 1995, RNA.

[18]  G. Beckett,et al.  Thyroidal extracellular glutathione peroxidase: a potential regulator of thyroid-hormone synthesis. , 1995, The Biochemical journal.

[19]  J. Köhrle,et al.  Differential selenium-dependent expression of type I 5'-deiodinase and glutathione peroxidase in the porcine epithelial kidney cell line LLC-PK1. , 1995, The Biochemical journal.

[20]  É. Fekete,et al.  Rapid stimulation of type I 5′-deiodinase in rat pituitaries by 3,3′,5-triiodo-l-thyronine , 1995, Molecular and Cellular Endocrinology.

[21]  T. Spelsberg,et al.  Development and characterization of a conditionally immortalized human fetal osteoblastic cell line , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[22]  R. Burk,et al.  Selenoprotein P. A selenium-rich extracellular glycoprotein. , 1994, The Journal of nutrition.

[23]  T. Yasushi,et al.  The human plasma glutathione peroxidase-encoding gene: organization, sequence and localization to chromosome 5q32 , 1994 .

[24]  M. Karnovsky,et al.  NADPH-oxidase expression and in situ production of superoxide by osteoclasts actively resorbing bone , 1994, The Journal of cell biology.

[25]  W. Ries,et al.  Superoxide and bone resorption. , 1994, Bone.

[26]  H. Blöcker,et al.  Phospholipid-hydroperoxide glutathione peroxidase. Genomic DNA, cDNA, and deduced amino acid sequence. , 1994, The Journal of biological chemistry.

[27]  M. Berry,et al.  Recognition of UGA as a selenocysteine codon in eukaryotes: a review of recent progress. , 1993, Biochemical Society transactions.

[28]  O. Jensen,et al.  Purification and properties of selenoprotein W from rat muscle. , 1993, The Journal of biological chemistry.

[29]  P. Newburger,et al.  Selenium regulation of glutathione peroxidase in human hepatoma cell line Hep3B. , 1993, Archives of biochemistry and biophysics.

[30]  J. Doroshow,et al.  Expression, characterization, and tissue distribution of a new cellular selenium-dependent glutathione peroxidase, GSHPx-GI. , 1993, The Journal of biological chemistry.

[31]  H. Notbohm,et al.  Fulvic acid supplementation and selenium deficiency disturb the structural integrity of mouse skeletal tissue. An animal model to study the molecular defects of Kashin-Beck disease. , 1993, The Biochemical journal.

[32]  R. Lloyd,et al.  Conserved nucleotide sequences in the open reading frame and 3' untranslated region of selenoprotein P mRNA. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Doroshow,et al.  Expression of plasma glutathione peroxidase in human liver in addition to kidney, heart, lung, and breast in humans and rodents. , 1992, Blood.

[34]  K. Cowan,et al.  Structure and function of the 5'-flanking sequence of the human cytosolic selenium-dependent glutathione peroxidase gene (hgpx1). , 1992, The Journal of biological chemistry.

[35]  J. Doroshow,et al.  Phospholipid hydroperoxide glutathione peroxidase is the 18-kDa selenoprotein expressed in human tumor cell lines. , 1991, The Journal of biological chemistry.

[36]  Kazuhiko Takahashi,et al.  Primary Structure of Human Plasma Glutathione Peroxidase Deduced from eDNA Sequences , 1990 .

[37]  L. Bonewald,et al.  Oxygen-derived free radicals stimulate osteoclastic bone resorption in rodent bone in vitro and in vivo. , 1990, The Journal of clinical investigation.

[38]  P. Newburger,et al.  Post-transcriptional regulation of glutathione peroxidase gene expression by selenium in the HL-60 human myeloid cell line. , 1989, Blood.

[39]  J. Whitin,et al.  Plasma selenium-dependent glutathione peroxidase. Cell of origin and secretion. , 1989, The Journal of biological chemistry.

[40]  Y. Ho,et al.  Nucleotide sequence of a rat glutathione peroxidase cDNA. , 1988, Nucleic acids research.

[41]  J. Whitin,et al.  Purification and characterization of human plasma glutathione peroxidase: a selenoglycoprotein distinct from the known cellular enzyme. , 1987, Archives of biochemistry and biophysics.

[42]  L. Marnett,et al.  Characterization of the hydroperoxide-reducing activity of human plasma. , 1987, Archives of biochemistry and biophysics.

[43]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[44]  A. Holmgren,et al.  Rat liver thioredoxin and thioredoxin reductase: purification and characterization. , 1982, Biochemistry.

[45]  J. L. Leonard,et al.  Iodothyronine 5'-deiodinase from rat kidney: substrate specificity and the 5'-deiodination of reverse triiodothyronine. , 1980, Endocrinology.

[46]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[47]  L. Flohé,et al.  Glutathione peroxidase: A selenoenzyme , 1973, FEBS letters.

[48]  H. Ganther,et al.  Selenium: Biochemical Role as a Component of Glutathione Peroxidase , 1973, Science.

[49]  K. Schwarz,et al.  Selenium as an integral part of Factor 3 against dietary necrotic liver degeneration. , 1957 .

[50]  P. Schleyer.  A SIMPLE PREPARATION OF ADAMANTANE , 1957 .

[51]  B. Otten,et al.  GH and TSH deficiency [letter] , 1998 .

[52]  G. Gamble,et al.  Evidence for secular change in Paget's disease. , 1997, Bone.

[53]  J. Köhrle,et al.  Expression of selenoproteins in various rat and human tissues and cell lines. , 1997, Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements.

[54]  M. Leist,et al.  Conventional cell culture media do not adequately supply cells with antioxidants and thus facilitate peroxide-induced genotoxicity. , 1996, Free radical biology & medicine.

[55]  D. Schomburg,et al.  Diversity of glutathione peroxidases. , 1995, Methods in enzymology.

[56]  M. Leist,et al.  Utilization of selenium from different chemical entities for selenoprotein biosynthesis by mammalian cell lines. , 1995, BioFactors.

[57]  A. Kyriakopoulos,et al.  Studies on the distribution and characteristics of new mammalian selenium-containing proteins. , 1995, The Analyst.

[58]  R. Burk.  Selenium in Biology and Human Health , 1994, Springer New York.

[59]  L. Key,et al.  Oxygen derived free radicals in osteoclasts: the specificity and location of the nitroblue tetrazolium reaction. , 1990, Bone.

[60]  Ernest Beutler,et al.  Red Cell Metabolism: A Manual of Biochemical Methods , 1975 .

[61]  P. Aiginger,et al.  [Clinical possibilities in the use of radioimmunologic TSH determination]. , 1974, Acta medica Austriaca.