Dual function of the selenoprotein PHGPx during sperm maturation

Science

Volumn 285, Issue 5432, 1999, Pages 1393-1396

  Ursini, Fulvio ^a   Heim, Sabina ^b   Kiess, Michael ^b   Maiorino, Matilde ^a   Roveri, Antonella ^a   Wissing, Josef ^b   Flohé, Leopold ^c  

^a UNIVERSITY OF PADOVA   (Italy)

^b HELMHOLTZ CENTRE FOR INFECTION RESEARCH   (Germany)

^c TECHNICAL UNIVERSITY OF BRAUNSCHWEIG   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

GLUTATHIONE PEROXIDASE; PHOSPHOLIPID DERIVATIVE; SELENOPROTEIN;

ARTICLE; CROSS LINKING; ENZYME ACTIVITY; HUMAN; MALE FERTILITY; MITOCHONDRION; NONHUMAN; OXIDATION; PRIORITY JOURNAL; SELENIUM DEFICIENCY; SPERMATID; SPERMATOZOON MATURATION;

ANIMALS; ELECTROPHORESIS, GEL, TWO-DIMENSIONAL; ELECTROPHORESIS, POLYACRYLAMIDE GEL; GLUTATHIONE PEROXIDASE; INFERTILITY, MALE; MALE; MITOCHONDRIA; OXIDATION-REDUCTION; PROTEINS; RATS; RATS, WISTAR; SELENIUM; SELENOPROTEINS; SOLUBILITY; SPECTROMETRY, MASS, MATRIX-ASSISTED LASER DESORPTION-IONIZATION; SPERMATIDS; SPERMATOGENESIS; SPERMATOZOA;

EID: 0033610030     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5432.1393     Document Type: Article

References (45)

1. D. G. Brown and R. F. Burk, J. Nutr. 103, 102 (1973); A. S. H. Wu, J. E. Oldfield, L. R. Shull, P. R. Cheeke, Biol. Reprod. 20, 793 (1979); E. Wallace, H. I. Calvin, K. Ploetz, G. W. Cooper, in Selenium in Biology and Medicine, G. F. Combs, O. A. Levander, J. E. Spallholz, J. E. Oldfield, Eds. (AVI, Westport, CT, 1987), pp. 181-196; D. Behne, H. Weiler, A. Kyriakopoulos, J. Reprod. Fertil. 106, 291 (1996); National Research Council Subcommittee on Selenium, Selenium in Nutrition (National Academy Press, Washington, DC, 1983).
2. D. G. Brown and R. F. Burk, J. Nutr. 103, 102 (1973); A. S. H. Wu, J. E. Oldfield, L. R. Shull, P. R. Cheeke, Biol. Reprod. 20, 793 (1979); E. Wallace, H. I. Calvin, K. Ploetz, G. W. Cooper, in Selenium in Biology and Medicine, G. F. Combs, O. A. Levander, J. E. Spallholz, J. E. Oldfield, Eds. (AVI, Westport, CT, 1987), pp. 181-196; D. Behne, H. Weiler, A. Kyriakopoulos, J. Reprod. Fertil. 106, 291 (1996); National Research Council Subcommittee on Selenium, Selenium in Nutrition (National Academy Press, Washington, DC, 1983).
3. D. G. Brown and R. F. Burk, J. Nutr. 103, 102 (1973); A. S. H. Wu, J. E. Oldfield, L. R. Shull, P. R. Cheeke, Biol. Reprod. 20, 793 (1979); E. Wallace, H. I. Calvin, K. Ploetz, G. W. Cooper, in Selenium in Biology and Medicine, G. F. Combs, O. A. Levander, J. E. Spallholz, J. E. Oldfield, Eds. (AVI, Westport, CT, 1987), pp. 181-196; D. Behne, H. Weiler, A. Kyriakopoulos, J. Reprod. Fertil. 106, 291 (1996); National Research Council Subcommittee on Selenium, Selenium in Nutrition (National Academy Press, Washington, DC, 1983).
4. D. G. Brown and R. F. Burk, J. Nutr. 103, 102 (1973); A. S. H. Wu, J. E. Oldfield, L. R. Shull, P. R. Cheeke, Biol. Reprod. 20, 793 (1979); E. Wallace, H. I. Calvin, K. Ploetz, G. W. Cooper, in Selenium in Biology and Medicine, G. F. Combs, O. A. Levander, J. E. Spallholz, J. E. Oldfield, Eds. (AVI, Westport, CT, 1987), pp. 181-196; D. Behne, H. Weiler, A. Kyriakopoulos, J. Reprod. Fertil. 106, 291 (1996); National Research Council Subcommittee on Selenium, Selenium in Nutrition (National Academy Press, Washington, DC, 1983).
5. D. G. Brown and R. F. Burk, J. Nutr. 103, 102 (1973); A. S. H. Wu, J. E. Oldfield, L. R. Shull, P. R. Cheeke, Biol. Reprod. 20, 793 (1979); E. Wallace, H. I. Calvin, K. Ploetz, G. W. Cooper, in Selenium in Biology and Medicine, G. F. Combs, O. A. Levander, J. E. Spallholz, J. E. Oldfield, Eds. (AVI, Westport, CT, 1987), pp. 181-196; D. Behne, H. Weiler, A. Kyriakopoulos, J. Reprod. Fertil. 106, 291 (1996); National Research Council Subcommittee on Selenium, Selenium in Nutrition (National Academy Press, Washington, DC, 1983).
6. J. T. Rotruck et al., Science 179, 588 (1973); L. Flohé, W. A. Günzler, H. H. Schock, FEBS Lett. 32, 132 (1973); D. C. Turner and T. C. Stadtman, Arch. Biochem. Biophys. 154, 366 (1973); J. R. Andreesen and L. G. Ljungdahl, J. Bacteriol. 116, 867 (1973).
7. J. T. Rotruck et al., Science 179, 588 (1973); L. Flohé, W. A. Günzler, H. H. Schock, FEBS Lett. 32, 132 (1973); D. C. Turner and T. C. Stadtman, Arch. Biochem. Biophys. 154, 366 (1973); J. R. Andreesen and L. G. Ljungdahl, J. Bacteriol. 116, 867 (1973).
8. J. T. Rotruck et al., Science 179, 588 (1973); L. Flohé, W. A. Günzler, H. H. Schock, FEBS Lett. 32, 132 (1973); D. C. Turner and T. C. Stadtman, Arch. Biochem. Biophys. 154, 366 (1973); J. R. Andreesen and L. G. Ljungdahl, J. Bacteriol. 116, 867 (1973).
9. J. T. Rotruck et al., Science 179, 588 (1973); L. Flohé, W. A. Günzler, H. H. Schock, FEBS Lett. 32, 132 (1973); D. C. Turner and T. C. Stadtman, Arch. Biochem. Biophys. 154, 366 (1973); J. R. Andreesen and L. G. Ljungdahl, J. Bacteriol. 116, 867 (1973).
10. A. Roveri et al., J. Biol. Chem. 267, 6142 (1992); A. Giannattasio, M. Girotti, K. Williams, L. Hall, A. J. Bellastella, J. Endocrinol. Invest. 20, 439 (1997); M. Maiorino et al., FASEB J. 12, 1359 (1998).
11. A. Roveri et al., J. Biol. Chem. 267, 6142 (1992); A. Giannattasio, M. Girotti, K. Williams, L. Hall, A. J. Bellastella, J. Endocrinol. Invest. 20, 439 (1997); M. Maiorino et al., FASEB J. 12, 1359 (1998).
12. A. Roveri et al., J. Biol. Chem. 267, 6142 (1992); A. Giannattasio, M. Girotti, K. Williams, L. Hall, A. J. Bellastella, J. Endocrinol. Invest. 20, 439 (1997); M. Maiorino et al., FASEB J. 12, 1359 (1998).
13. H. I. Calvin, G. W. Cooper, E. Wallace, Gamete Res. 4, 139 (1981).
14. L. Cataldo, K. Baig, R. Oko, M. A. Mastrangelo, K. C. Kleene, Mol. Reprod. Dev. 45, 320 (1996).
15. V. Pallini and E. Bacci, J. Submicr. Cytol. 11, 165 (1979); S.-Y. Nam, H.-Y. Youn, K. Ogawa, M. Kurohmaru, Y. Hayashi, J. Reprod. Dev. 43, 227 (1997).
16. V. Pallini and E. Bacci, J. Submicr. Cytol. 11, 165 (1979); S.-Y. Nam, H.-Y. Youn, K. Ogawa, M. Kurohmaru, Y. Hayashi, J. Reprod. Dev. 43, 227 (1997).
17. I. M. Adham et al., DNA Cell Biol. 15, 159 (1996).
18. T. C. Stadtman, Annu. Rev. Biochem. 59, 111 (1990); J. Heider, Ch. Baron, A. Böck, EMBO J. 11, 3759 (1992); M. J. Berry, L. Banu, J. W. Harney, P. R. Larsen, ibid. 12, 3315 (1993); L. Flohé, E. Wingender, R. Brigelius-Flohé, in Oxidative Stress and Signal Transduction, H. J. Forman and E. Cadenas, Eds. (Chapman & Hall, New York, 1997), pp. 415-440.
19. T. C. Stadtman, Annu. Rev. Biochem. 59, 111 (1990); J. Heider, Ch. Baron, A. Böck, EMBO J. 11, 3759 (1992); M. J. Berry, L. Banu, J. W. Harney, P. R. Larsen, ibid. 12, 3315 (1993); L. Flohé, E. Wingender, R. Brigelius-Flohé, in Oxidative Stress and Signal Transduction, H. J. Forman and E. Cadenas, Eds. (Chapman & Hall, New York, 1997), pp. 415-440.
20. T. C. Stadtman, Annu. Rev. Biochem. 59, 111 (1990); J. Heider, Ch. Baron, A. Böck, EMBO J. 11, 3759 (1992); M. J. Berry, L. Banu, J. W. Harney, P. R. Larsen, ibid. 12, 3315 (1993); L. Flohé, E. Wingender, R. Brigelius-Flohé, in Oxidative Stress and Signal Transduction, H. J. Forman and E. Cadenas, Eds. (Chapman & Hall, New York, 1997), pp. 415-440.
21. T. C. Stadtman, Annu. Rev. Biochem. 59, 111 (1990); J. Heider, Ch. Baron, A. Böck, EMBO J. 11, 3759 (1992); M. J. Berry, L. Banu, J. W. Harney, P. R. Larsen, ibid. 12, 3315 (1993); L. Flohé, E. Wingender, R. Brigelius-Flohé, in Oxidative Stress and Signal Transduction, H. J. Forman and E. Cadenas, Eds. (Chapman & Hall, New York, 1997), pp. 415-440.
22. Spermatozoa of 4-month-old Wistar rats were collected by squeezing the cauda epididymis and vas deferens in phosphate-buffered saline (PBS) and centrifuged at 600g for 10 min. Spermatogenic cells were prepared as described [M. L. Meistrich, J. Longtin, W. A. Brock, S. R. Grimes, M. L. Mace, Biol. Reprod. 25, 1065 (1981)]. Sperm mitochondrial capsule was prepared according to (4). 10. Proteins were blotted onto nitrocellulose, probed with an antigen-purified rabbit antibody raised against pig heart PHGPx, and detected by biotinylated antibody to rabbit immunogtobulin G (IgG) and streptavidin alkaline phosphatase complex.
23. 2-terminal sequencing, proteins were blotted onto polyvinylidene difluoride membranes for 16 hours at pH 8.3 (25 mM tris-HCl, 192 mW glycine) and 100 mA (30 V).
24. Mitochondrial capsule material (100 μg) was dissolved in 400 μl of a solution containing 7 M urea, 2 M thiourea, 4% CHAPS, 40 mM DTT, 20 mM tris base, and 0.5% IPG buffer (Pharmacia) and focused in an IPG-phor (Pharmacia) at 20°C by stepwise Increasing voltage up to 5000 V at <30 μA per IPG strip. The pH gradient was nonlinear from 3 to 10 or linear from 3 to 10 or 6 to 11. The focused IPG strips were equilibrated for SDS-PAGE with a solution containing 60 mM DTT in 6 M urea, 30% glycerol and 0.05 M tris-HCl buffer (pH 8.8), and thereafter with the same buffer containing 250 mM iodoacetamide instead of DTT. Gels were stained with Coomassie blue.
25. 3 overnight at 37°C. The digests were extracted twice, dried, and reconstituted in 10 μl of water. Peptides were separated on a reversed-phase capillary column (0.5 mm × 150 mm) with a gradient of acetonitrile in 0.1% formic acid/4 mM ammonium acetate at a flow rate of 5 μl/min. Aliquots of 5 μl were spotted onto Biobrene-treated glass fiber filters and sequenced on an Applied Biosystems 494A sequencer with standard pulsed liquid cycles.
26. + ion from two peptide standards; 100 to 200 laser shots were summed for each spectrum. Mass identification was performed with MS-Fit (http://falcon.ludwig.ucl.ac.uk/ucsfhtml3.2/msfit. htm).
27. 2-terminus of the size isomers starting with Ala-Ser-Arg-Asp-Asp-Trp-Arg-Cys-Ala-Arg, a sequence either corresponding to the originally proposed translation start [R. Brigelius-Flohé et al., J. Biol. Chem. 269, 7342 (1994)] after cleavage of the first two residues or derived from pre-PHGPx [T. R. Pushpa-Rekha, L. M. Burdsal, G. M. Chisolm, D. M. Driscoll, ibid. 270, 26993 (1995)] after processing of the mitochondrial leader peptide [M. Arai et al., Biochem. Biophys. Res. Commun. 227, 433 (1996)]. Tryptic fragments extending toward the COOH-terminus up to position 164 were also observed with the faster migrating specimen. Charge heterogeneity may arise from phosphorylation [R. Schuckelt et al., Free Radical Res. Commun. 14, 343 (1991)], deaminations of Gln and Asn residues, COOH-terminal degradation, and oxidation or elimination of selenium.
28. 2-terminus of the size isomers starting with Ala-Ser-Arg-Asp-Asp- Trp-Arg-Cys-Ala-Arg, a sequence either corresponding to the originally proposed translation start [R. Brigelius-Flohé et al., J. Biol. Chem. 269, 7342 (1994)] after cleavage of the first two residues or derived from pre-PHGPx [T. R. Pushpa-Rekha, L. M. Burdsal, G. M. Chisolm, D. M. Driscoll, ibid. 270, 26993 (1995)] after processing of the mitochondrial leader peptide [M. Arai et al., Biochem. Biophys. Res. Commun. 227, 433 (1996)]. Tryptic fragments extending toward the COOH-terminus up to position 164 were also observed with the faster migrating specimen. Charge heterogeneity may arise from phosphorylation [R. Schuckelt et al., Free Radical Res. Commun. 14, 343 (1991)], deaminations of Gln and Asn residues, COOH-terminal degradation, and oxidation or elimination of selenium.
29. 2-terminus of the size isomers starting with Ala-Ser-Arg-Asp-Asp- Trp-Arg-Cys-Ala-Arg, a sequence either corresponding to the originally proposed translation start [R. Brigelius-Flohé et al., J. Biol. Chem. 269, 7342 (1994)] after cleavage of the first two residues or derived from pre-PHGPx [T. R. Pushpa-Rekha, L. M. Burdsal, G. M. Chisolm, D. M. Driscoll, ibid. 270, 26993 (1995)] after processing of the mitochondrial leader peptide [M. Arai et al., Biochem. Biophys. Res. Commun. 227, 433 (1996)]. Tryptic fragments extending toward the COOH-terminus up to position 164 were also observed with the faster migrating specimen. Charge heterogeneity may arise from phosphorylation [R. Schuckelt et al., Free Radical Res. Commun. 14, 343 (1991)], deaminations of Gln and Asn residues, COOH-terminal degradation, and oxidation or elimination of selenium.
30. 2-terminus of the size isomers starting with Ala-Ser-Arg-Asp-Asp- Trp-Arg-Cys-Ala-Arg, a sequence either corresponding to the originally proposed translation start [R. Brigelius-Flohé et al., J. Biol. Chem. 269, 7342 (1994)] after cleavage of the first two residues or derived from pre-PHGPx [T. R. Pushpa-Rekha, L. M. Burdsal, G. M. Chisolm, D. M. Driscoll, ibid. 270, 26993 (1995)] after processing of the mitochondrial leader peptide [M. Arai et al., Biochem. Biophys. Res. Commun. 227, 433 (1996)]. Tryptic fragments extending toward the COOH-terminus up to position 164 were also observed with the faster migrating specimen. Charge heterogeneity may arise from phosphorylation [R. Schuckelt et al., Free Radical Res. Commun. 14, 343 (1991)], deaminations of Gln and Asn residues, COOH-terminal degradation, and oxidation or elimination of selenium.
31. A. Roveri, M. Maiorino, F. Ursini, Methods Enzymol. 233, 202 (1994).
32. Solubilization and reduction were carried out in 0.1 M tris-HCl, 6 M guanidine-HCl, pepstatin A (0.5 μg/ml), leupeptin (0.7 μg/ml), and 0.1 M 2-mercaptoethanol at pH 7.5 and 4°C for 10 min. After centrifugation (150,000g for 30 min), low molecular weight compounds were removed by a NAP 5 cartridge equilibrated with 0.01 M tris-HCl, 0.15 M NaCl, 1 mM EDTA, and 1% Triton X-100 at pH 7.5. PHGPx activity was measured at 37°C with phosphatidylcholine hydroperoxide at 3 mM CSH according to (16). Control samples were treated identically but with 5 mM 2-mercaptoethanol.
33. Other examples of "gene sharing" or "moonlighting proteins," respectively, are reviewed by J. Piatigorsky, Prog. Ret. Eye Res. 17, 145 (1998); C. J. Jefferey, Trends Biol. Sci. 24, 8 (1999).
34. Other examples of "gene sharing" or "moonlighting proteins," respectively, are reviewed by J. Piatigorsky, Prog. Ret. Eye Res. 17, 145 (1998); C. J. Jefferey, Trends Biol. Sci. 24, 8 (1999).
35. M. Maiorino et al., Biol. Chem. Hoppe Seyler 376, 651 (1995); F. Ursini et al., Methods Enzymol. 252, 38 (1995).
36. M. Maiorino et al., Biol. Chem. Hoppe Seyler 376, 651 (1995); F. Ursini et al., Methods Enzymol. 252, 38 (1995).
37. F. Bauché, M.-H. Fouchard, B. Jégou, FEBS Lett. 349, 392 (1994); R. Shalgi, J. Seligman, N. S. Kosower, Biol. Reprod. 40, 1037 (1989); J. Seligman, N. S. Kosower, R. Shalgi, ibid. 46, 301 (1992); H. M. Fisher and R. J. Aitken, J. Exp. Zool. 277, 390 (1997).
38. F. Bauché, M.-H. Fouchard, B. Jégou, FEBS Lett. 349, 392 (1994); R. Shalgi, J. Seligman, N. S. Kosower, Biol. Reprod. 40, 1037 (1989); J. Seligman, N. S. Kosower, R. Shalgi, ibid. 46, 301 (1992); H. M. Fisher and R. J. Aitken, J. Exp. Zool. 277, 390 (1997).
39. F. Bauché, M.-H. Fouchard, B. Jégou, FEBS Lett. 349, 392 (1994); R. Shalgi, J. Seligman, N. S. Kosower, Biol. Reprod. 40, 1037 (1989); J. Seligman, N. S. Kosower, R. Shalgi, ibid. 46, 301 (1992); H. M. Fisher and R. J. Aitken, J. Exp. Zool. 277, 390 (1997).
40. F. Bauché, M.-H. Fouchard, B. Jégou, FEBS Lett. 349, 392 (1994); R. Shalgi, J. Seligman, N. S. Kosower, Biol. Reprod. 40, 1037 (1989); J. Seligman, N. S. Kosower, R. Shalgi, ibid. 46, 301 (1992); H. M. Fisher and R. J. Aitken, J. Exp. Zool. 277, 390 (1997).
41. F. Weitzel and A. Wendel, J. Biol. Chem. 268, 6288 (1993); R. Brigelius-Flohé, B. Friedrichs, S. Maurer, M. Schultz, R. Streicher, Biochem. J. 328, 199 (1997); P. A. Sandstrom, J. Murray, T. M. Folks, A. M. Diamond, Free Radical Biol. Med. 24, 1485 (1998).
42. F. Weitzel and A. Wendel, J. Biol. Chem. 268, 6288 (1993); R. Brigelius-Flohé, B. Friedrichs, S. Maurer, M. Schultz, R. Streicher, Biochem. J. 328, 199 (1997); P. A. Sandstrom, J. Murray, T. M. Folks, A. M. Diamond, Free Radical Biol. Med. 24, 1485 (1998).
43. F. Weitzel and A. Wendel, J. Biol. Chem. 268, 6288 (1993); R. Brigelius-Flohé, B. Friedrichs, S. Maurer, M. Schultz, R. Streicher, Biochem. J. 328, 199 (1997); P. A. Sandstrom, J. Murray, T. M. Folks, A. M. Diamond, Free Radical Biol. Med. 24, 1485 (1998).
44. In other gels, mitochondrial glutathione S-transferase subunit Yb-2 (accession number 121719) and endothelin converting enzyme (NCBI accession number 1706564) could be identified by MALDI-TOF or peptide sequencing.
45. Supported by the German Ministry of Education, Science and Technology, the Italian Ministry of University and Scientific Research, National Research Council, Italy, and the BIOMED 2 program of the European Community.