Biology Reference
In-Depth Information
CHAPTER
2
Pierre Germain
*
,{
and William Bourguet
*
,{
Dimerization of Nuclear
Receptors
*
Inserm U1054, Centre de Biochimie Structurale, Montpellier, France
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CNRS UMR5048, Universite´s Montpellier 1 & 2, Montpellier, France
CHAPTER OUTLINE
Introduction .............................................................................................................. 22
2.1 Methods ............................................................................................................ 24
2.1.1 Studies of NR-NR Interactions through Protein Crystallization ............. 24
2.1.1.1 Required Materials .....................................................................26
2.1.1.2 Protocol......................................................................................26
2.1.2 Expression and Purification of NR-NR Complexes ............................... 27
2.1.2.1 Required Materials .....................................................................28
2.1.2.2 Protocol......................................................................................29
2.1.3 Monitoring NR-NR Interactions by Noncovalent Electrospray Ionization
Mass Spectrometry ........................................................................... 29
2.1.3.1 Required Materials .....................................................................30
2.1.3.2 Protocol......................................................................................30
2.1.4 Monitoring NR-NR Interactions by Electrophoretic Mobility Shift
Assays .......................................................................................... 31
2.1.4.1 Required Materials .....................................................................31
2.1.4.2 Protocol......................................................................................31
2.1.5 Two-hybrid Assays to Define NR-NR Interactions in Living Cells........... 32
2.1.5.1 Required Materials .....................................................................34
2.1.5.2 Protocol......................................................................................34
2.1.6 Fluorescence Cross-Correlation Spectroscopy to Measure the
Concentrations and Interactions of NRs in Living Cells ........................ 35
2.1.6.1 Required Materials .....................................................................36
2.1.6.2 Protocol......................................................................................36
Acknowledgments ..................................................................................................... 39
References ............................................................................................................... 39
