Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics

Kate Watt, Iain J McEwan

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy have proved powerful methods for studying the structure of the isolated ligand and DNA-binding domains of nuclear receptors. However, the N-terminal domain (NTD), which in some members of the superfamily is important for transcriptional regulation, and the full-length receptor proteins have proved more challenging. The NTD of different nuclear receptors show little sequence homology and can vary dramatically in length from a few to several hundred amino acids. Low resolution structural analysis using circular dichroism, NMR, steady-state fluorescence spectroscopy, and Fourier transformed infrared spectroscopy has provided valuable information on the conformation and folding of the structurally plastic NTD. In this chapter, we discuss protocols for measuring the intrinsic fluorescence emission spectrum for tryptophan residues under different experimental conditions of protein folding and unfolding.
Original languageEnglish
Title of host publicationThe Nuclear Receptor Superfamily
Subtitle of host publicationMethods and Protocols
EditorsIain J McEwan
PublisherHumana Press
Pages205-218
Number of pages14
ISBN (Electronic)978-1-60327-575-0
ISBN (Print)978-1-60327-574-3
DOIs
Publication statusPublished - 2009

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Volume505
ISSN (Print)1064-3745

Fingerprint

Steroid Receptors
Fluorescence spectroscopy
Emission spectroscopy
Cytoplasmic and Nuclear Receptors
Conformations
Protein folding
X ray crystallography
Structural analysis
Tryptophan
Nuclear magnetic resonance spectroscopy
Infrared spectroscopy
Proteins
Fluorescence
Nuclear magnetic resonance
Plastics
Ligands
Amino Acids
DNA

Cite this

Watt, K., & McEwan, I. J. (2009). Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics. In I. J. McEwan (Ed.), The Nuclear Receptor Superfamily: Methods and Protocols (pp. 205-218). (Methods in Molecular Biology; Vol. 505). Humana Press. https://doi.org/10.1007/978-1-60327-575-0_12

Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics. / Watt, Kate; McEwan, Iain J.

The Nuclear Receptor Superfamily: Methods and Protocols. ed. / Iain J McEwan. Humana Press, 2009. p. 205-218 (Methods in Molecular Biology; Vol. 505).

Research output: Chapter in Book/Report/Conference proceedingChapter

Watt, K & McEwan, IJ 2009, Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics. in IJ McEwan (ed.), The Nuclear Receptor Superfamily: Methods and Protocols. Methods in Molecular Biology, vol. 505, Humana Press, pp. 205-218. https://doi.org/10.1007/978-1-60327-575-0_12
Watt K, McEwan IJ. Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics. In McEwan IJ, editor, The Nuclear Receptor Superfamily: Methods and Protocols. Humana Press. 2009. p. 205-218. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-60327-575-0_12
Watt, Kate ; McEwan, Iain J. / Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics. The Nuclear Receptor Superfamily: Methods and Protocols. editor / Iain J McEwan. Humana Press, 2009. pp. 205-218 (Methods in Molecular Biology).
@inbook{6634dee7ee05455eb9393f80a4789d2a,
title = "Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics",
abstract = "X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy have proved powerful methods for studying the structure of the isolated ligand and DNA-binding domains of nuclear receptors. However, the N-terminal domain (NTD), which in some members of the superfamily is important for transcriptional regulation, and the full-length receptor proteins have proved more challenging. The NTD of different nuclear receptors show little sequence homology and can vary dramatically in length from a few to several hundred amino acids. Low resolution structural analysis using circular dichroism, NMR, steady-state fluorescence spectroscopy, and Fourier transformed infrared spectroscopy has provided valuable information on the conformation and folding of the structurally plastic NTD. In this chapter, we discuss protocols for measuring the intrinsic fluorescence emission spectrum for tryptophan residues under different experimental conditions of protein folding and unfolding.",
author = "Kate Watt and McEwan, {Iain J}",
year = "2009",
doi = "10.1007/978-1-60327-575-0_12",
language = "English",
isbn = "978-1-60327-574-3",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "205--218",
editor = "McEwan, {Iain J}",
booktitle = "The Nuclear Receptor Superfamily",

}

TY - CHAP

T1 - Using intrinsic fluorescence emission spectroscopy to study steroid receptor and coactivator protein conformation dynamics

AU - Watt, Kate

AU - McEwan, Iain J

PY - 2009

Y1 - 2009

N2 - X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy have proved powerful methods for studying the structure of the isolated ligand and DNA-binding domains of nuclear receptors. However, the N-terminal domain (NTD), which in some members of the superfamily is important for transcriptional regulation, and the full-length receptor proteins have proved more challenging. The NTD of different nuclear receptors show little sequence homology and can vary dramatically in length from a few to several hundred amino acids. Low resolution structural analysis using circular dichroism, NMR, steady-state fluorescence spectroscopy, and Fourier transformed infrared spectroscopy has provided valuable information on the conformation and folding of the structurally plastic NTD. In this chapter, we discuss protocols for measuring the intrinsic fluorescence emission spectrum for tryptophan residues under different experimental conditions of protein folding and unfolding.

AB - X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy have proved powerful methods for studying the structure of the isolated ligand and DNA-binding domains of nuclear receptors. However, the N-terminal domain (NTD), which in some members of the superfamily is important for transcriptional regulation, and the full-length receptor proteins have proved more challenging. The NTD of different nuclear receptors show little sequence homology and can vary dramatically in length from a few to several hundred amino acids. Low resolution structural analysis using circular dichroism, NMR, steady-state fluorescence spectroscopy, and Fourier transformed infrared spectroscopy has provided valuable information on the conformation and folding of the structurally plastic NTD. In this chapter, we discuss protocols for measuring the intrinsic fluorescence emission spectrum for tryptophan residues under different experimental conditions of protein folding and unfolding.

U2 - 10.1007/978-1-60327-575-0_12

DO - 10.1007/978-1-60327-575-0_12

M3 - Chapter

SN - 978-1-60327-574-3

T3 - Methods in Molecular Biology

SP - 205

EP - 218

BT - The Nuclear Receptor Superfamily

A2 - McEwan, Iain J

PB - Humana Press

ER -