Abstract
Crystallographic analysis of the ligand-binding domains of nuclear hormone receptors (NR) has provided a unique insight into the molecular events that underlie the ligand-mediated control of their transcriptional activity. The technique relies on preparing milligram quantities of protein, growing three-dimensional crystals of the desired protein—ligand complex, collecting X-ray diffraction data, and subsequently interpreting the derived electron density maps to reveal the structure of the complex.
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References
Pike, A. C. W. (2006) Lessons learnt from structural studies of the oestrogen receptor. Best Practice&Research Clinical Endocrinology&Metabolism. 20, 1–14.
Li, Y., Lambert, M. H., Xu, H. E. (2003) Activation of nuclear receptors: A perspective from structural genomics. Structure. 11, 741–746.
Bergfors, T. (1999) Protein Crystallization:Techniques, Strategies, and Tips. A Laboratory Manual. La Jolla, USA: International University Line.
McRee, D. E. (1999) Practical Protein Crystallography. 2nd ed. San Diego: Academic Press.
Blow, D. (2002) Outline of Crystallography for Biologists. Oxford: Oxford University Press.
Benvenuti, M., Mangani, S. (2007) Crystallization of soluble proteins in vapor diffusion for X-ray crystallography. Nature Protocols. 2, 1633–1651.
Carlson, K. E., Choi, I., Gee, A., Katzenel-lenbogen, B. S., Katzenellenbogen, J. A. (1997) Altered ligand binding properties and enhanced stability of a constitutively active estrogen receptor: Evidence that an open pocket conformation is required for ligand interaction. Biochemistry. 36, 14897–14905.
Miroux, B., Walker, J. E. (1996) Over-production of proteins in Escherichia coli: Mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. Journal of Molecular Biology. 260, 289–298.
Laemmli, U. K. (1970) Cleavage of structural proteins during assembly of head of bacteri-ophage-T4. Nature. 227, 680–685.
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Pike, A.C. (2009). X-Ray Crystallography of Agonist/Antagonist-Bound Receptors. In: McEwan, I.J. (eds) The Nuclear Receptor Superfamily. Methods in Molecular Biology™, vol 505. Humana Press. https://doi.org/10.1007/978-1-60327-575-0_4
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DOI: https://doi.org/10.1007/978-1-60327-575-0_4
Publisher Name: Humana Press
Print ISBN: 978-1-60327-574-3
Online ISBN: 978-1-60327-575-0
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