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Analysis of Ethylene Receptors: Ethylene-Binding Assays

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Ethylene Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1573))

Abstract

Plant ethylene receptors bind ethylene with high affinity. Most of the characterization of ethylene binding to the receptors has been carried out using a radioligand-binding assay on functional receptors expressed in yeast. In this chapter, we describe methods for expressing ethylene receptors in yeast and conducting ethylene-binding assays on intact yeast and yeast membranes. The ethylene-binding assays can be modified to analyze ethylene binding to intact plants and other organisms as well as membranes isolated from any biological source.

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References

  1. Mattoo AK, Suttle JC (eds) (1991) The Plant Hormone Ethylene. CRC Press, Inc., Boca Raton

    Google Scholar 

  2. Abeles FB, Morgan PW, M. E. Saltveit J (1992) Ethylene in Plant Biology. 2nd edn. Academic Press, San Diego

    Google Scholar 

  3. Neljubov D (1901) Uber die horizontale nutation der Stengel von Pisum sativum und einiger anderer. Pflanzen Beih Bot Zentralb 10:128–139

    Google Scholar 

  4. Sisler EC (1979) Measurement of ethylene binding in plant tissue. Plant Physiol 64:538–542

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Sisler EC (1980) Partial purification of an ethylene-binding component from plant tissue. Plant Physiol 66:404–406

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Evans DE, Bengochea T, Cairns AJ, Dodds JH, Hall MA (1982) Studies on ethylene binding by cell-free preparations from cotyledons of Phaseolus vulgaris L.: subcellular localization. Plant Cell Environ 5:101–107

    CAS  Google Scholar 

  7. Evans DE, Dodds JH, Lloyd PC, apGwynn I, Hall MA (1982) A study of the subcellular localisation of an ethylene binding site in developing cotyledons of Phaseolus vulgaris L. by high resolution autoradiography. Planta 154:48-52.

    Google Scholar 

  8. Bleecker AB, Estelle MA, Somerville C, Kende H (1988) Insensitivity to ethylene conferred by a dominant mutation in Arabidopsis thaliana. Science 241:1086–1089

    Article  CAS  PubMed  Google Scholar 

  9. Guzmán P, Ecker JR (1990) Exploiting the triple response of Arabidopsis to identify ethylene-related mutants. Plant Cell 2:513–523

    Article  PubMed  PubMed Central  Google Scholar 

  10. Chang C, Kwok SF, Bleecker AB, Meyerowitz EM (1993) Arabidopsis ethylene-response gene ETR1: similarity of product to two-component regulators. Science 262(5133):539–544

    Article  CAS  PubMed  Google Scholar 

  11. Schaller GE, Bleecker AB (1995) Ethylene-binding sites generated in yeast expressing the Arabidopsis ETR1 gene. Science 270(5243):1809–1811

    Article  CAS  PubMed  Google Scholar 

  12. Schaller GE, Ladd AN, Lanahan MB, Spanbauer JM, Bleecker AB (1995) The ethylene response mediator ETR1 from Arabidopsis forms a disulfide-linked dimer. J Biol Chem 270(21):12526–12530

    Article  CAS  PubMed  Google Scholar 

  13. Rodriguez FI, Esch JJ, Hall AE, Binder BM, Schaller GE, Bleecker AB (1999) A copper cofactor for the ethylene receptor ETR1 from Arabidopsis. Science 283(5404):996–998

    Article  CAS  PubMed  Google Scholar 

  14. Fink GR, Guthrie C (eds) (2002) Methods in enzymology: guide to yeast genetics and molecular cell biology, Part B, vol 350. Academic Press, San Diego, CA

    Google Scholar 

  15. Amberg DC, Burck D, Strathern JN (eds) (2005) Methods in yeast genetics: a cold spring harbor laboratory course manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  16. McDaniel B, Binder BM (2012) Ethylene receptor1 (ETR1) is sufficient and has the predominant role in mediating inhibition of ethylene responses by silver in Arabidopsis thaliana. J Biol Chem 287:26094–26103. doi:10.1074/jbc.M112.383034

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Binder BM, Rodriguez FI, Bleecker AB, Patterson SE (2007) The effects of group 11 transition metals, including gold, on ethylene binding to the ETR1 receptor and growth of Arabidopsis thaliana. FEBS Lett 581:5105–5109

    Article  CAS  PubMed  Google Scholar 

  18. Wang W, Esch JE, Shiu SH, Agula H, Binder BM, Chang C, Patterson SE, Bleecker AB (2006) Identification of important regions for ethylene binding and signaling in the transmembrane domain of the ETR1 ethylene receptor of Arabidopsis. Plant Cell 18:3429–3442

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Blankenship SM, Sisler EC (1993) Ethylene binding site affinity in ripening apples. J Am Soc Hort Sci 118(5):609–612

    CAS  Google Scholar 

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Acknowledgments

This work was supported by NSF grants IOS-1456487 (to GES), IOS-1254423 (to BMB), and MCB-1517032 (to GES and BMB).

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Authors and Affiliations

  1. Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, M407 Walters Life Sciences, 1414 Cumberland Ave., Knoxville, TN, 37996, USA

    Brad M. Binder

  2. Department of Biological Sciences, Dartmouth College, Hanover, NH, 03755, USA

    G. Eric Schaller

Authors
  1. Brad M. Binder
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  2. G. Eric Schaller
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Corresponding author

Correspondence to Brad M. Binder .

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Editors and Affiliations

  1. Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee, USA

    Brad M. Binder

  2. Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire, USA

    G. Eric Schaller

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Binder, B.M., Schaller, G.E. (2017). Analysis of Ethylene Receptors: Ethylene-Binding Assays. In: Binder, B., Eric Schaller, G. (eds) Ethylene Signaling. Methods in Molecular Biology, vol 1573. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6854-1_7

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  • DOI: https://doi.org/10.1007/978-1-4939-6854-1_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6852-7

  • Online ISBN: 978-1-4939-6854-1

  • eBook Packages: Springer Protocols

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