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Cytotechnology

, Volume 57, Issue 2, pp 145–150 | Cite as

Development of an in vitro system for screening the ligands of a membrane glycoprotein CD36

  • Hitomi Inagaki
  • Satoshi Tsuzuki
  • Takashi Iino
  • Kazuo Inoue
  • Tohru Fushiki
JAACT Special Issue.

Abstract

It has well been known that human and rodents exhibit a preference for fats. This suggests the existence of an orosensory system responsible for the detection of dietary fats. A plasma membrane glycoprotein CD36, besides the role in the uptake of long-chain fatty acids (LCFAs) as well as oxidized low-density lipoprotein (OxLDL) in a variety of cells, has been postulated to be a candidate fat taste receptor on the tongue. Therefore, molecules that bind with CD36 to cause intracellular signaling but have fewer calories could be substitutes for dietary fats. In the present study, we developed an in vitro system for the screening of CD36 ligands using Chinese hamster ovary-K1 cells (CHO-K1) stably transfected with human or mouse CD36. When incubated with OxLDL labeled with fluorescence dye, the fluorescence was much higher in the transfected CHO-K1 cells than in non-transfected CHO-K1 cells. Incubation of the transfected cells with OxLDL caused a rapid phosphorylation of extracellular signal regulated kinase, and the degree was significantly higher compared with that in non-transfected CHO-K1 cells. The expression system using CHO-K1 cells could be a convenient tool to screen the novel ligands of CD36.

Keywords

CD36 Ligands Oxidized LDL Non-oxidized LDL Binding ERK1/2 

Notes

Acknowledgments

This study has been supported by Program for Promotion of Basic Research Activities for Innovative Bioscience.

References

  1. Abumrad NA, el-Maghrabi MR, Amri EZ et al (1993) Cloning of a rat adipocyte membraneprotein implicated in binding or transport of long-chain fatty acids that is induced during preadipocyte differentiation. Homology with human CD36. J Biol Chem 268:17665–17668Google Scholar
  2. Boullier A, Gillotte KL, Horkko S et al (2000) The binding of oxidized low density lipoprotein to mouse CD36 is mediated in part by oxidized phospholipids that are associated with both the lipid and protein moieties of the lipoprotein. J Biol Chem 275:9163–9169CrossRefGoogle Scholar
  3. Brown MS, Goldstein JL (1979) Receptor-mediated endocytosis: insights from the lipoprotein receptor system. Proc Natl Acad Sci USA 76:3330–3337CrossRefGoogle Scholar
  4. Febbraio M, Hajjar DP, Silverstein RL (2001) CD36: a class B scavenger receptor involved in angiogenesis, atherosclerosis, inflammation, and lipid metabolism. J Clin Invest 108:785–791Google Scholar
  5. Fukuwatari T, Kawada T, Tsuruta M et al (1997) Expression of the putative membrane fatty acid transporter (FAT) in taste buds of the circumvallate papillae in rats. FEBS Lett 414:461–464CrossRefGoogle Scholar
  6. Greenwalt DE, Lipsky RH, Ockenhouse CF et al (1992) Membrane glycoprotein CD36: a review of its roles in adherence, signal transduction, and transfusion medicine. Blood 80:1105–1115Google Scholar
  7. Hill CS, Treisman R (1995) Transcriptional regulation by extracellular signals: mechanisms and specificity. Cell 80:199–211CrossRefGoogle Scholar
  8. Kyriakis JM, Avruch J (2001) Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 81:807–869Google Scholar
  9. Laugerette F, Passilly-Degrace P, Patris B et al (2005) CD36 involvement in orosensory detection of dietary lipids, spontaneous fat preference, and digestive secretions. J Clin Invest 115:3177–3184CrossRefGoogle Scholar
  10. Marshall CJ (1995) Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80:179–185CrossRefGoogle Scholar
  11. Pearce SF, Roy P, Nicholson AC et al (1998) Recombinant glutathione S-transferase/CD36 fusion proteins define an oxidized low density lipoprotein-binding domain. J Biol Chem 273:34875–34881CrossRefGoogle Scholar
  12. Tabata T, Mine S, Kawahara C, Okada Y et al (2002) Monocyte chemoattractant protein-1 induces scavenger receptor expression and monocyte differentiation into foam cells. APMIS 110:458–468CrossRefGoogle Scholar
  13. Tandon NN, Kralisz U, Jamieson GA (1989) Identification of glycoprotein IV (CD36) as a primary receptor for platelet-collagen adhesion. J Biol Chem 264:7576–7583Google Scholar
  14. Tibbles LA, Woodgett JR (1999) The stress-activated protein kinase pathways. Cell Mol Life Sci 55:1230–1254CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Hitomi Inagaki
    • 1
  • Satoshi Tsuzuki
    • 1
  • Takashi Iino
    • 2
  • Kazuo Inoue
    • 1
  • Tohru Fushiki
    • 1
  1. 1.Laboratory of Nutrition Chemistry, Division of Food Science and Biotechnology, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Pharmafoods International Co. Ltd.KyotoJapan

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