scholarly journals Spatial Intensity Distribution Analysis: Studies of G Protein-Coupled Receptor Oligomerisation

2018 ◽  
Vol 39 (2) ◽  
pp. 175-186 ◽  
Author(s):  
John D. Pediani ◽  
Richard J. Ward ◽  
Sara Marsango ◽  
Graeme Milligan
Keyword(s):  
G Protein ◽  
Intensity Distribution ◽  
Distribution Analysis ◽  
G Protein Coupled Receptor ◽  
Spatial Intensity Distribution ◽  
Spatial Intensity ◽  
G Protein Coupled

scholarly journals Spatial intensity distribution analysis quantifies the extent and regulation of homodimerization of the secretin receptor

2017 ◽  
Vol 474 (11) ◽  
pp. 1879-1895 ◽  
Author(s):  
Richard J. Ward ◽  
John D. Pediani ◽  
Kaleeckal G. Harikumar ◽  
Laurence J. Miller ◽  
Graeme Milligan
Keyword(s):  
G Protein ◽  
Intensity Distribution ◽  
Transmembrane Domain ◽  
Distribution Analysis ◽  
Wild Type ◽  
Spatial Intensity Distribution ◽  
Spatial Intensity ◽  
Secretin Receptor ◽  
Dimeric Form ◽  
Type Receptor

Previous studies have indicated that the G-protein-coupled secretin receptor is present as a homodimer, organized through symmetrical contacts in transmembrane domain IV, and that receptor dimerization is critical for high-potency signalling by secretin. However, whether all of the receptor exists in the dimeric form or if this is regulated is unclear. We used measures of quantal brightness of the secretin receptor tagged with monomeric enhanced green fluorescent protein (mEGFP) and spatial intensity distribution analysis to assess this. Calibration using cells expressing plasma membrane-anchored forms of mEGFP initially allowed us to demonstrate that the epidermal growth factor receptor is predominantly monomeric in the absence of ligand and while wild-type receptor was rapidly converted into a dimeric form by ligand, a mutated form of this receptor remained monomeric. Equivalent studies showed that, at moderate expression levels, the secretin receptor exists as a mixture of monomeric and dimeric forms, with little evidence of higher-order complexity. However, sodium butyrate-induced up-regulation of the receptor resulted in a shift from monomeric towards oligomeric organization. In contrast, a form of the secretin receptor containing a pair of mutations on the lipid-facing side of transmembrane domain IV was almost entirely monomeric. Down-regulation of the secretin receptor-interacting G-protein Gαs did not alter receptor organization, indicating that dimerization is defined specifically by direct protein–protein interactions between copies of the receptor polypeptide, while short-term treatment with secretin had no effect on organization of the wild-type receptor but increased the dimeric proportion of the mutated receptor variant.

Spatial intensity distribution Analysis (SpIDA): A fluorescence microscopy based method to measure receptor oligomerization in cells

2012 ◽  
Vol 18 (S2) ◽  
pp. 128-129 ◽  
Author(s):  
P.W. Wiseman ◽  
A.G. Godin ◽  
J.L. Swift ◽  
S. Costantino ◽  
A. Ribeiro-da-Silva ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Intensity Distribution ◽  
Distribution Analysis ◽  
Spatial Intensity Distribution ◽  
Spatial Intensity ◽  
Receptor Oligomerization ◽  
In Cells

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Distribution analysis of nonsynonymous polymorphisms within the G-protein-coupled receptor gene family

Genomics ◽  
2003 ◽  
Vol 81 (3) ◽  
pp. 245-248 ◽  
Author(s):  
Andria Lee ◽  
B.K. Rana ◽  
H.H. Schiffer ◽  
N.J. Schork ◽  
M.R. Brann ◽  
...  
Keyword(s):  
Gene Family ◽  
G Protein ◽  
Receptor Gene ◽  
Distribution Analysis ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled ◽  
Receptor Gene Family

Spatial Intensity Distribution Analysis (SpIDA)

2013 ◽  
pp. 1-19 ◽  
Author(s):  
Annie Barbeau ◽  
Jody L. Swift ◽  
Antoine G. Godin ◽  
Yves De Koninck ◽  
Paul W. Wiseman ◽  
...  
Keyword(s):  
Intensity Distribution ◽  
Distribution Analysis ◽  
Spatial Intensity Distribution ◽  
Spatial Intensity

scholarly journals Revealing protein oligomerization and densities in situ using spatial intensity distribution analysis

2011 ◽  
Vol 108 (17) ◽  
pp. 7010-7015 ◽  
Author(s):  
A. G. Godin ◽  
S. Costantino ◽  
L.-E. Lorenzo ◽  
J. L. Swift ◽  
M. Sergeev ◽  
...  
Keyword(s):  
Intensity Distribution ◽  
Protein Oligomerization ◽  
Distribution Analysis ◽  
Spatial Intensity Distribution ◽  
Spatial Intensity
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