Functional analysis of G-protein coupled receptors using a new fluorescein lactone-based intracellular calcium indicator

2010 ◽  
Vol 2 (3) ◽  
pp. 295 ◽  
Author(s):  
Desuo Yang ◽  
Chunmei Wei ◽  
Jinfang Liao ◽  
Zhenjun Diwu
Keyword(s):  
Functional Analysis ◽  
Intracellular Calcium ◽  
G Protein ◽  
G Protein Coupled Receptors ◽  
Calcium Indicator ◽  
G Protein Coupled

scholarly journals Identification of a Novel Family of Targets of PYK2 Related to Drosophila Retinal Degeneration B (rdgB) Protein

1999 ◽  
Vol 19 (3) ◽  
pp. 2278-2288 ◽  
Author(s):  
Sima Lev ◽  
John Hernandez ◽  
Ricardo Martinez ◽  
Alon Chen ◽  
Greg Plowman ◽  
...  
Keyword(s):  
Intracellular Calcium ◽  
Retinal Degeneration ◽  
G Protein ◽  
Binding Proteins ◽  
G Protein Coupled Receptors ◽  
Sequence Motif ◽  
Phosphoinositide Metabolism ◽  
Amino Terminal ◽  
Terminal Domain ◽  
G Protein Coupled

ABSTRACT The protein tyrosine kinase PYK2 has been implicated in signaling pathways activated by G-protein-coupled receptors, intracellular calcium, and stress signals. Here we describe the molecular cloning and characterization of a novel family of PYK2-binding proteins designated Nirs (PYK2 N-terminal domain-interacting receptors). The three Nir proteins (Nir1, Nir2, and Nir3) bind to the amino-terminal domain of PYK2 via a conserved sequence motif located in the carboxy terminus. The primary structures of Nirs reveal six putative transmembrane domains, a region homologous to phosphatidylinositol (PI) transfer protein, and an acidic domain. The Nir proteins are the human homologues of the Drosophila retinal degeneration B protein (rdgB), a protein implicated in the visual transduction pathway in flies. We demonstrate that Nirs are calcium-binding proteins that exhibit PI transfer activity in vivo. Activation of PYK2 by agents that elevate intracellular calcium or by phorbol ester induce tyrosine phosphorylation of Nirs. Moreover, PYK2 and Nirs exhibit similar expression patterns in several regions of the brain and retina. In addition, PYK2-Nir complexes are detected in lysates prepared from cultured cells or from brain tissues. Finally, the Nir1-encoding gene is located at human chromosome 17p13.1, in proximity to a locus responsible for several human retinal diseases. We propose that the Nir and rdgB proteins represent a new family of evolutionarily conserved PYK2-binding proteins that play a role in the control of calcium and phosphoinositide metabolism downstream of G-protein-coupled receptors.

An atlas and functional analysis of G-protein coupled receptors in human islets of Langerhans

2013 ◽  
Vol 139 (3) ◽  
pp. 359-391 ◽  
Author(s):  
Stefan Amisten ◽  
Albert Salehi ◽  
Patrik Rorsman ◽  
Peter M. Jones ◽  
Shanta J. Persaud
Keyword(s):  
Functional Analysis ◽  
G Protein ◽  
Islets Of Langerhans ◽  
Human Islets ◽  
G Protein Coupled Receptors ◽  
Human Islets Of Langerhans ◽  
G Protein Coupled

scholarly journals Lysophosphatidylcholine Containing Anisic Acid Is Able to Stimulate Insulin Secretion Targeting G Protein Coupled Receptors

Nutrients ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 1173 ◽  
Author(s):  
Anna Drzazga ◽  
Marta Okulus ◽  
Magdalena Rychlicka ◽  
Łukasz Biegała ◽  
Anna Gliszczyńska ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Insulin Secretion ◽  
Intracellular Calcium ◽  
G Protein ◽  
Phenolic Acids ◽  
G Protein Coupled Receptors ◽  
Calcium Flux ◽  
Pancreatic Cell ◽  
Anisic Acid ◽  
G Protein Coupled

Diabetes mellitus is a worldwide health problem with high rates of mortality and morbidity. Management of diabetes mellitus by dietary components is achievable especially at the initial stage of the disease. Several studies confirmed the antidiabetic activities of simple phenolic acids and lysophosphatidylcholine (LPC). The main goal of this study was to identify new potential insulin secretion modulators obtained by combining the structures of two natural compounds, namely O-methyl derivatives of phenolic acids and phospholipids. LPC and phosphatidylcholine bearing methoxylated aromatic carboxylic acids were tested as potential agents able to improve glucose-stimulated insulin secretion (GSIS) and intracellular calcium mobilization in MIN6 β pancreatic cell line. Our results show that LPC with covalently bonded molecule of p-anisic acid at the sn-1 position was able to induce GSIS and intracellular calcium flux. Notably, 1-anisoyl-2-hydroxy-sn-glycero-3-phosphocholine did not affect the viability of MIN6 cells, suggesting its potential safe use. Furthermore, we have shown that three G protein coupled receptors, namely GPR40, GPR55, and GPR119, are targeted by this LPC derivative.

scholarly journals Evaluation of FLIPR Calcium 3 Assay Kit—A New No-Wash Fluorescence Calcium Indicator Reagent

2003 ◽  
Vol 8 (5) ◽  
pp. 571-577 ◽  
Author(s):  
Yingxin Zhang ◽  
Dianne Kowal ◽  
Angela Kramer ◽  
John Dunlop
Keyword(s):  
Calcium Signaling ◽  
G Protein ◽  
Glutamate Receptor ◽  
Metabotropic Glutamate Receptor ◽  
G Protein Coupled Receptors ◽  
Metabotropic Glutamate Receptor 5 ◽  
Metabotropic Glutamate ◽  
Calcium Indicator ◽  
Indicator Dye ◽  
G Protein Coupled

We have evaluated the FLIPR Calcium 3 Assay Kit (Calcium 3), a new no-wash fluorescence calcium indicator dye reagent, for the measurement of agonist-stimulated calcium signaling in cells expressing the serotonin 2C (5-HT2C), metabotropic glutamate receptor 5 (mGluR5) and the vasopressin 2 (V2) G-protein-coupled receptors. Calcium 3 yielded equivalent (5-HT2C) or superior (mGluR5 and V2) sensitivity to FLUO-4 as indexed by the change in fluorescence counts following agonist application. Assay variability, indexed by CV, using Calcium 3 or FLUO-4 was equivalent with 5-HT2C receptor responses although CVs were reduced using Calcium 3 in the examples of the mGluR5 and V2 receptors. Receptor pharmacologies based on agonist EC50 values were identical when either Calcium 3 or FLUO-4 were utilized. Our results validate Calcium 3 as a compel-ling alternative to FLUO-4 in the choice of fluorescent dye reagent for studying G-protein-coupled receptors, providing the advantage of a homogenous, no-wash assay format. ( Journal of Biomolecular Screening 2003:571-577)

Functional Screening of G Protein–Coupled Receptors by Measuring Intracellular Calcium with a Fluorescence Microplate Reader

2002 ◽  
Vol 7 (3) ◽  
pp. 233-246 ◽  
Author(s):  
Matthias U. Kassack ◽  
Barbara Höfgen ◽  
Jochen Lehmann ◽  
Niels Eckstein ◽  
J. Mark Quillan ◽  
...  
Keyword(s):  
Intracellular Calcium ◽  
G Protein ◽  
G Protein Coupled Receptors ◽  
Functional Screening ◽  
Microplate Reader ◽  
G Protein Coupled
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