GPCR Oligomerization: Contribution to Receptor Biogenesis - GPCR Signalling Complexes: Synthesis, Assembly, Trafficking and Specificity - page 42

Chapter 3
GPCR Oligomerization: Contribution
to Receptor Biogenesis

Kathleen Van Craenenbroeck

Contents
3.1 Introduction..................................................................................................................... 44

3.2 How to Study Oligomerization During Biogenesis ........................................................ 45

3.2.1 Co-immunoprecipitation...................................................................................... 46

3.2.2 Biophysical Methods ........................................................................................... 47

3.2.2.1 Fluorescence Resonance Energy Transfer

Techniques (FRET)............................................................................... 49

3.2.2.2 Bioluminescence Resonance Energy Transfer

Techniques (BRET) .............................................................................. 50

3.2.2.3 Bimolecular Fluorescence or Luminescence

Complementation (BiFC/BiLC) ........................................................... 51

3.2.3 Proximity Ligation Assay (PLA)......................................................................... 51

3.2.4 The Use of Mutants: Dominant Negative and ER Trapping Studies................... 52

3.3 Folding of GPCRs........................................................................................................... 53

3.3.1 Translation of GPCR mRNA............................................................................... 53

3.3.2 Receptor Biogenesis in the Endoplasmic Reticulum........................................... 54

3.4 Exit from the ER and Trafficking to the Plasma Membrane .......................................... 56

3.5 Functionality of GPCRs at the Plasma Membrane ......................................................... 56

3.6 Role of Dimerization in the Biogenesis of Different

GPCR Family Members.................................................................................................. 57

3.6.1 Family A .............................................................................................................. 57

3.6.2 Family B .............................................................................................................. 58

3.6.3 Family C .............................................................................................................. 58

3.7 Receptor-Receptor Interaction Domains: The GPCR
as a Chaperone, Promoting Folding................................................................................ 59
3.8 How Biogenesis of GPCR Oligomers can be Influenced ............................................... 60
3.8.1 Role of the Availability of Different GPCRs....................................................... 60
3.8.2 Availability of GPCR Ligands............................................................................. 61
3.9 Concluding Remarks ...................................................................................................... 61
References ................................................................................................................................ 61