scholarly journals Discrepancy between fluorescence correlation spectroscopy and fluorescence recovery after photobleaching diffusion measurements of G-protein-coupled receptors

2013 ◽  
Vol 440 (1) ◽  
pp. 40-48 ◽  
Author(s):  
Rhodora Cristina Calizo ◽  
Suzanne Scarlata
Keyword(s):  
G Protein ◽  
Fluorescence Correlation Spectroscopy ◽  
Fluorescence Recovery After Photobleaching ◽  
Correlation Spectroscopy ◽  
G Protein Coupled Receptors ◽  
Fluorescence Recovery ◽  
Fluorescence Correlation ◽  
G Protein Coupled

scholarly journals The use of fluorescence correlation spectroscopy to characterize the molecular mobility of fluorescently labelled G protein-coupled receptors

2016 ◽  
Vol 44 (2) ◽  
pp. 624-629 ◽  
Author(s):  
Laura E. Kilpatrick ◽  
Stephen J. Hill
Keyword(s):  
G Protein ◽  
Fluorescence Correlation Spectroscopy ◽  
Photon Counting ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
G Protein Coupled Receptors ◽  
Fluorescence Correlation ◽  
Membrane Bound ◽  
Molecular Brightness ◽  
G Protein Coupled

The membranes of living cells have been shown to be highly organized into distinct microdomains, which has spatial and temporal consequences for the interaction of membrane bound receptors and their signalling partners as complexes. Fluorescence correlation spectroscopy (FCS) is a technique with single cell sensitivity that sheds light on the molecular dynamics of fluorescently labelled receptors, ligands or signalling complexes within small plasma membrane regions of living cells. This review provides an overview of the use of FCS to probe the real time quantification of the diffusion and concentration of G protein-coupled receptors (GPCRs), primarily to gain insights into ligand–receptor interactions and the molecular composition of signalling complexes. In addition we document the use of photon counting histogram (PCH) analysis to investigate how changes in molecular brightness (ε) can be a sensitive indicator of changes in molecular mass of fluorescently labelled moieties.

The use of fluorescence correlation spectroscopy to characterise the molecular mobility of G protein-coupled receptors in membrane microdomains: an update

2021 ◽  
Vol 49 (4) ◽  
pp. 1547-1554
Author(s):  
Laura E. Kilpatrick ◽  
Stephen J. Hill
Keyword(s):  
G Protein ◽  
Fluorescence Correlation Spectroscopy ◽  
Molecular Mechanisms ◽  
Short Review ◽  
Correlation Spectroscopy ◽  
G Protein Coupled Receptors ◽  
Membrane Microdomains ◽  
Sensitivity Threshold ◽  
Fluorescence Correlation ◽  
G Protein Coupled

It has become increasingly apparent that some G protein-coupled receptors (GPCRs) are not homogeneously expressed within the plasma membrane but may instead be organised within distinct signalling microdomains. These microdomains localise GPCRs in close proximity with other membrane proteins and intracellular signalling partners and could have profound implications for the spatial and temporal control of downstream signalling. In order to probe the molecular mechanisms that govern GPCR pharmacology within these domains, fluorescence techniques with effective single receptor sensitivity are required. Of these, fluorescence correlation spectroscopy (FCS) is a technique that meets this sensitivity threshold. This short review will provide an update of the recent uses of FCS based techniques in conjunction with GPCR subtype selective fluorescent ligands to characterise dynamic ligand–receptor interactions in whole cells and using purified GPCRs.

scholarly journals Single molecule binding of a ligand to a G-protein-coupled receptor in real time using fluorescence correlation spectroscopy, rendered possible by nano-encapsulation in styrene maleic acid lipid particles

Nanoscale ◽  
2020 ◽  
Vol 12 (21) ◽  
pp. 11518-11525 ◽  
Author(s):  
Rachael L. Grime ◽  
Joelle Goulding ◽  
Romez Uddin ◽  
Leigh A. Stoddart ◽  
Stephen J. Hill ◽  
...  
Keyword(s):  
Real Time ◽  
G Protein ◽  
Single Molecule ◽  
Maleic Acid ◽  
Fluorescence Correlation Spectroscopy ◽  
Correlation Spectroscopy ◽  
G Protein Coupled Receptor ◽  
Fluorescence Correlation ◽  
Lipid Particles ◽  
G Protein Coupled

Combining the technologies of encapsulation of GPCRs in SMA lipid particles with fluorescence correlation spectroscopy provides a versatile characterisation platform.

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