BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
권호 표지
DOI QR코드

DOI QR Code

A Simple ELISA for Screening Ligands of Peroxisome Proliferator-activated Receptor γ

  • Cho, Min-Chul (Laboratory of Cellular Biology, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Hae-Sook (Laboratory of Cellular Biology, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Jae-Hwa (Laboratory of Cellular Biology, Korea Research Institute of Bioscience and Biotechnology) ;
  • Choe, Yong-Kyung (Laboratory of Cellular Biology, Korea Research Institute of Bioscience and Biotechnology) ;
  • Hong, Jin-Tae (College of Pharmacy, Chungbuk National University) ;
  • Paik, Sang-Gi (Department of Biology, Chungnam National University) ;
  • Yoon, Do-Young (Laboratory of Cellular Biology, Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2003.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Peroxisome proliferator-activated receptors (PPARs) are orphan nuclear hormone receptors that are known to control the expression of genes that are involved in lipid homeostasis and energy balance. PPARs activate gene transcription in response to a variety of compounds, including hypolipidemic drugs. Most of these compounds have high affinity to the ligand-binding domain (LBD) of PPARs and cause a conformational change within PPARs. As a result, the receptor is converted to an activated mode that promotes the recruitment fo co-activators such as the steroid receptor co-activator-1 (SRC-1). Based on the activation mechanism of PPARs (the ligand binding to $PPAR{\gamma}$ induces interactions of the receptor with transcriptional co-activators), we performed Western blot and ELISA. These showed that the indomethacin, a $PPAR{\gamma}$ ligand, increased the binding between $PPAR{\gamma}$ and SRC-1 in a ligand dose-dependent manner. These results suggested that the in vitro conformational change of $PPAR{\gamma}$ by ligands was also induced, and increased the levels of the ligand-dependent interaction with SRC-1. Collectively, we developed a novel and useful ELISA system for the mass screening of $PPAR{\gamma}$ ligands. This screening system (based on the interaction between $PPAR{\gamma}$ and SRC-1) may be a promising system in the development of drugs for metabolic disorders.

Keywords

References

  1. Baumann, C. A., Chokshi, N., Saltiel, A. R. and Ribon, V. (2000) Cloning and characterization of a functional peroxisome proliferator activator receptor-gamma-responsive element in the promoter of the CAP gene. J. Biol. Chem. 275, 9131-9135. https://doi.org/10.1074/jbc.275.13.9131
  2. Chen, S., Johnson, B. A., Li, Y., Aster, S., McKeever, B., Mosley, R., Moller, D. E. and Zhou, G. (2000) Both co-activator LXXLL motif-dependent and -independent interactions are required for peroxisome proliferator-activated receptor gamma (PPARgamma) function. J. BioI. Chem. 275, 3733-3736. https://doi.org/10.1074/jbc.275.6.3733
  3. Cho, Y. S., Cho, C. W., Kang, J. W., Cho, M. C., Lee, K. A., Shim, J. H., Kwon, D. H., Choe, Y. K., Park, S. N. and Yoon, D. Y. (2001) Development of a screening system for drugs against human papillomavirus-associated cervical cancer: Based on E7-Rb binding. J. Biochem. Mol. Biol. 34, 80-84.
  4. Durand, B., Saunders, M., Gaudon, C., Roy, B., Lasson, R. and Chambon, P. (1994) Activation function 2 (AF-2) of retinoic acid receptor and 9-cis retinoic acid receptor: presence of a conserved autonomous constitutive activating domain and influence of the nature of the response element on AF-2 activity. EMBO. J. 13, 5370-5382.
  5. Elbrecht, A., Chen, Y., Adams, A., Berger, J., Griffin, P., Klatt, T., Zhang, B., Menke, J., Zhou, G., Smith, R. G. and Moller, D. E. (1999) L-764406 is a partial agonist of human peroxisome proliferator-activated receptor. J. Biol. Chem. 274, 7913-7922. https://doi.org/10.1074/jbc.274.12.7913
  6. Elstner, E., Muller, C., Koshizuka, K, Williamson, E. A., Park, D., Asou, H., Shintaku, P., Said, J, W., Heber, D. and Koeffler, H. P. (1998) Ligands for peroxisome proliferator-activated receptorgamma and retinoic acid receptor inhibit growth and induce apoptosis of human breast cancer cells in vitro and in BNX mice. Proc. Natl. Acad. Sci. USA 95, 8806-8811. https://doi.org/10.1073/pnas.95.15.8806
  7. Forman, B. M., Chen, J. and Evans, R. M. (1997) Hypolipidemic drugs, polyunsaturated fatty acids, and eicosanoids are ligands for peroxisome proliferator-activated receptors alpha and delta. Proc. Natl. Acad. Sci. USA 94, 4312-4317. https://doi.org/10.1073/pnas.94.9.4312
  8. Giguere, V. (1999) Orphan Nuclear Receptors: From Gene to Function. Endocr. Rev. 20, 689-725. https://doi.org/10.1210/er.20.5.689
  9. Hertz, R., Berman, I., Keppler, D. and Bar-Tana, J. (1996) Activation of gene transcription by prostacyclin analogues is mediated by the peroxisome-proliferators-activated receptor (PPAR). Eur. J. Biochem. 235, 242-247. https://doi.org/10.1111/j.1432-1033.1996.00242.x
  10. Horwitz, K. B., Jackson, T. A., Bain, D. L., Richer, J. K, Takimoto, G. S. and Tung, L. (1996) Nuclear receptor co- activators and co-repressors. Mol. Endocrinol. 10, 1167-77. https://doi.org/10.1210/me.10.10.1167
  11. Issemann, I., Prince, R. A., Tugwood, J. D. and Green, S. (1993) The peroxisome proliferator-activated receptor:retinoid X receptor heterodimer is activated by fatty acids and fibrate hypolipidaemic drugs. J. Mol. Endocrinol. 11, 37-47. https://doi.org/10.1677/jme.0.0110037
  12. Krey, G., Braissant, O., L'Horset, F., Kalkhoven, E., Perroud, M., Parker, M. G. and Wahli, W. (1997) Fatty acids, eicosanoids, and hypolipidemic agents identified as ligands of peroxisome proliferator-activated receptors by co-activator-dependent receptor ligand assay. Mol. Endocrinol. 11, 779-791. https://doi.org/10.1210/me.11.6.779
  13. Lee, I. H., Kim, I. S. and Lee, S. K. (2000) Expression of all- trans retinoic acid glycoprotein and inflammatory cytokines during differentiation of HL-60 cell. J. Biochem. Mol. Biol. 33, 402-406.
  14. Lee, S. S., Lee, K. L., Yoo, J. Y., Jeong, M. J., Park, Y. M., Kwon, B. M., Bae, Y. S. and Han, M. Y. (2001) Development of ELISA system for screening of specific binding inhibitors for Src homology(SH)2 domain and phosphotyrosine interactions. J. Biochem. Mol. Biol. 34, 537-543.
  15. Lehmann, J. M., Lenhard, J. M., Oliver, B. B., Ringold, G. M. and Kliewer, S. A. (1997) Peroxisome proliferator-activated receptors alpha and gamma are activated by indomethacin and other non-steroidal anti-inflammatory drugs. J. Biol. Chem. 272, 3406-3410. https://doi.org/10.1074/jbc.272.6.3406
  16. Park, J. S., Lee, H. G., Lee, Y., Kang, Y H., Rhim, H. S. and Choi, S. Y. (2000) Expression of human immunodeficiency virus type 1 Tat proteins in Escherichia coli and application to study Tat functions. J. Biochem. Mol. BioI. 33, 337-343.
  17. Sambrook, J. Fritsch, E. F. and Maniatis, T. (1989) Molecular cloning: A laboratory manual. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, New York.
  18. Schmidt, A., Endo, N., Rutledge, S. J., Vogel, R., Shinar, D. and Rodan, G. A. (1992) Identitication of a new member of the steroid hormone receptor superfamily that is activated by a peroxisome proliferator and fatty acids. Mol. Endocrinol. 6, 1634-1641. https://doi.org/10.1210/me.6.10.1634
  19. Shao, D., Rangwala, S. M., Bailey, S. T., Krakow, S. L., Reginato, M. J. and Lazar, M. A. (1998) Interdomain communication regulating ligand binding by PPAR-gamma. Nature 396, 377- 380. https://doi.org/10.1038/24634
  20. Tolon, R. M., Castillo, A. I. and Aranda, A. (1998) Activation of the prolactin gene by peroxisome proliferator-activated receptor- alpha appears to be DNA binding-independent. J. Biol. Chem. 273, 26652-26661. https://doi.org/10.1074/jbc.273.41.26652
  21. Tontonoz, P., Hu, E., Graves, R, A., Budavari, A. I. and Spiegelman, B. M. (1994) mPPAR gamma 2: tissue-specific regulator of an adipocyte enhancer. Genes Dev. 8, 1224-1234. https://doi.org/10.1101/gad.8.10.1224
  22. Tontonoz, P., Singer, S., Forman, B. M., Sarraf, P., Fletcher. J. A., Fletcher, C. D., Brun, R. P., Mueller, E., Altiok, S., Oppenheim, H., Evans, R. M. and Spiegelman, B. M. (1997) Tenninal differentiation of human liposarcoma cells induced by ligands for peroxisome proliferator-activated receptor gamma and the retinoid X receptor. Proc. Natl. Acad. Sci. USA 94, 237-241. https://doi.org/10.1073/pnas.94.1.237
  23. Torchia, J., Rose, D. W, Inostroza, J., Kamei, Y., Westin, S., Glass, C. K. and Rosenfeld, M. G. (1997) The transcriptional co-activator p/CIP binds CBP and mediates nuclear-receptor function. Nature 387, 677-684. https://doi.org/10.1038/42652
  24. Voegel, J. J., Heine, M. J., Zechel, C., Chambon, P. and Gronemeyer, H. (1996) TIF2, a 160 kDa transcriptional mediator for the ligand-dependent activation function AF-2 of nuclear receptors. EMBO. J. 15, 3667-3675.
  25. Werman, A., Hollenberg, A., Solanes, G., Bjorbaek, C., Vidal-Puig A. J. and Flier, J. S. (1997) Ligand-independent activation domain in the N terminus of peroxisome proliferator-activated receptor gamma (PPARgamma). J. BioI. Chem. 272, 20230-20235 https://doi.org/10.1074/jbc.272.32.20230
  26. Xin, X., Yang, S., Kowalski, J. and Gerritsen, M. E. (1999) Peroxisome proliferator-activated receptor gamma ligands are potent inhibitors of angiogenesis in vitro and in vivo. J. BioI. Chem. 274, 9116-9121. https://doi.org/10.1074/jbc.274.13.9116
  27. Yu, K., Bayona, W, Kallen, C. B., Harding, H. P., Ravera, C. P., McMahon, G., Brown, M. and Lazar, M. A. (1995) Differential activation of peroxisome proliferator-activated receptors by eicosanoids. J. Biol. Chem. 270, 23975-23983. https://doi.org/10.1074/jbc.270.41.23975
  28. Zhou, G., Cummings, R., Hermes, J. and Moller, D. E. (2001) Use of homogeneous time-resolved fluorescence energy transfer in the measurement of nuclear receptor activation. Methods 25, 54-61. https://doi.org/10.1006/meth.2001.1215
  29. Zhu, Y., Qi, C., Calandra, C., Rao, M. S. and Reddy, J. K. (1996) Cloning and identification of mouse steroid receptor coactivator-1 (mSRC-1), as a co-activator of peroxisome proliferator-activated receptor gamma. Gene Expr. 6, 185-195.

Cited by

  1. 5-(3,5-Di-tert-butyl-4-hydroxybenzylidene) thiazolidine-2,4-dione modulates peroxisome proliferators-activated receptor γ in 3T3-L1 adipocytes: Roles as a PPARγ ligand vol.242, pp.1-2, 2005, https://doi.org/10.1016/j.mce.2005.08.005
  2. A simple method to screen ligands of peroxisome proliferator-activated receptor δ vol.29, pp.5, 2006, https://doi.org/10.1016/j.ejps.2006.07.003
  3. Peroxisome Proliferators-Activated Receptor (PPAR) Modulators and Metabolic Disorders vol.2008, 2008, https://doi.org/10.1155/2008/679137
  4. Epitope analysis of PPARγ monoclonal antibody Pγ48.34A and its application for screening PPARγ ligands vol.296, pp.1-2, 2005, https://doi.org/10.1016/j.jim.2004.11.011
  5. IL-18 E42A mutant is resistant to the inhibitory effects of HPV-16 E6 and E7 oncogenes on the IL-18-mediated immune response vol.229, pp.2, 2005, https://doi.org/10.1016/j.canlet.2005.06.042
  6. A Role for the PPARγin Cancer Therapy vol.2008, 2008, https://doi.org/10.1155/2008/314974
  7. Platelet lysate suppresses the expression of lipocalin-type prostaglandin D2 synthase that positively controls adipogenic differentiation of human mesenchymal stromal cells vol.318, pp.18, 2012, https://doi.org/10.1016/j.yexcr.2012.06.004