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Analysis of Interaction of Estradiol with Estrogen Receptor by NMR Spectroscopy

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Estrogen Receptors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1366))

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Abstract

Following binding to estradiol, estrogen receptors (ER) α and ERβ recruit a number of interacting proteins and mediate a plethora of functions. The binding of estrogen with the receptors shows changes in the resonance structure and movement of protons. We cloned ERβ and its trans-activation domain (TAD) and ligand-binding domain (LBD), expressed them in prokaryotic expression vectors, purified them, and studied their interaction with estradiol. In this chapter, a detailed method of preparation of recombinant proteins, SDS-PAGE, silver staining, and NMR are described. Such methods are useful to check the biological activity of bacterially expressed proteins and are applicable to basic and applied research.

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References

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Acknowledgements

Financial support from UGC-CAS and Department of Biotechnology, Government of India, to M.K.T. is highly acknowledged.

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Authors and Affiliations

  1. Biochemistry and Molecular Biology Laboratory, Centre of Advanced Study, Department of Zoology, Banaras Hindu University, Varanasi, UP, 221 005, India

    M. K. Thakur & V. Paramanik

  2. Department of Zoology, Indira Gandhi National Tribal University, Kapil Dhara Road, Mekal Sadan, Amarkantak, Anuppur, MP, 484 887, India

    V. Paramanik

Authors
  1. M. K. Thakur
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  2. V. Paramanik
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Corresponding author

Correspondence to V. Paramanik .

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Editors and Affiliations

  1. University of South Dakota Sanford School of Medicine, Vermillion, South Dakota, USA

    Kathleen M. Eyster

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Thakur, M.K., Paramanik, V. (2016). Analysis of Interaction of Estradiol with Estrogen Receptor by NMR Spectroscopy. In: Eyster, K.M. (eds) Estrogen Receptors. Methods in Molecular Biology, vol 1366. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3127-9_18

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  • DOI: https://doi.org/10.1007/978-1-4939-3127-9_18

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3126-2

  • Online ISBN: 978-1-4939-3127-9

  • eBook Packages: Springer Protocols

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