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Kinetics and Dynamics in the G Protein-Coupled Receptor Signaling Cascade

We describe optical and microscopy methods based on Förster resonance energy transfer, fluorescence recovery after photobleaching, and imaging cross-correlation spectroscopy that permit to determine kinetic and dynamic properties of key reactions involved G protein-coupled receptor (GPCR) signaling... Full description

Journal Title: Methods in Enzymology 2013, Vol.522, p.337-363
Main Author: Vilardaga, Jean-Pierre
Other Authors: Romero, Guillermo , Feinstein, Timothy N , Wehbi, Vanessa L
Format: Electronic Article Electronic Article
Language: English
Subjects:
Quelle: Gale eBooks
Publisher: United States: Elsevier Science & Technology
ID: ISSN: 0076-6879
Link: https://www.ncbi.nlm.nih.gov/pubmed/23374192
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title: Kinetics and Dynamics in the G Protein-Coupled Receptor Signaling Cascade
format: Article
creator:
  • Vilardaga, Jean-Pierre
  • Romero, Guillermo
  • Feinstein, Timothy N
  • Wehbi, Vanessa L
subjects:
  • Arrestins - genetics
  • Arrestins - metabolism
  • Cyclic AMP - metabolism
  • Fluorescence
  • Fluorescence Resonance Energy Transfer - methods
  • FRAP
  • FRET
  • G protein-coupled receptors
  • G proteins
  • Gene Expression
  • HEK293 Cells
  • Heterotrimeric GTP-Binding Proteins - genetics
  • Heterotrimeric GTP-Binding Proteins - metabolism
  • Humans
  • ICCS
  • Kinetics
  • Ligands
  • Membrane proteins
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Parathyroid Hormone - metabolism
  • Parathyroid hormone receptor
  • Photobleaching
  • Protein Binding
  • Protein Stability
  • Receptors, Parathyroid Hormone - genetics
  • Receptors, Parathyroid Hormone - metabolism
  • Signal transduction
  • Signal Transduction - genetics
ispartof: Methods in Enzymology, 2013, Vol.522, p.337-363
description: We describe optical and microscopy methods based on Förster resonance energy transfer, fluorescence recovery after photobleaching, and imaging cross-correlation spectroscopy that permit to determine kinetic and dynamic properties of key reactions involved G protein-coupled receptor (GPCR) signaling from the initial ligand binding step to the generation of the second messenger, cAMP. Well suited to determine rate-limiting reactions taking place along a GPCR signaling cascade in live cells, these techniques have also uncovered new concepts in GPCR signaling as well as many interesting mechanistic subtleties by which GPCRs transmit neurotransmitter and hormone signals into cells.
language: eng
source: Gale eBooks
identifier: ISSN: 0076-6879
fulltext: fulltext
issn:
  • 0076-6879
  • 1557-7988
url: Link


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descriptionWe describe optical and microscopy methods based on Förster resonance energy transfer, fluorescence recovery after photobleaching, and imaging cross-correlation spectroscopy that permit to determine kinetic and dynamic properties of key reactions involved G protein-coupled receptor (GPCR) signaling from the initial ligand binding step to the generation of the second messenger, cAMP. Well suited to determine rate-limiting reactions taking place along a GPCR signaling cascade in live cells, these techniques have also uncovered new concepts in GPCR signaling as well as many interesting mechanistic subtleties by which GPCRs transmit neurotransmitter and hormone signals into cells.
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subjectArrestins - genetics ; Arrestins - metabolism ; Cyclic AMP - metabolism ; Fluorescence ; Fluorescence Resonance Energy Transfer - methods ; FRAP ; FRET ; G protein-coupled receptors ; G proteins ; Gene Expression ; HEK293 Cells ; Heterotrimeric GTP-Binding Proteins - genetics ; Heterotrimeric GTP-Binding Proteins - metabolism ; Humans ; ICCS ; Kinetics ; Ligands ; Membrane proteins ; Microscopy, Confocal ; Microscopy, Fluorescence ; Parathyroid Hormone - metabolism ; Parathyroid hormone receptor ; Photobleaching ; Protein Binding ; Protein Stability ; Receptors, Parathyroid Hormone - genetics ; Receptors, Parathyroid Hormone - metabolism ; Signal transduction ; Signal Transduction - genetics
ispartofMethods in Enzymology, 2013, Vol.522, p.337-363
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descriptionWe describe optical and microscopy methods based on Förster resonance energy transfer, fluorescence recovery after photobleaching, and imaging cross-correlation spectroscopy that permit to determine kinetic and dynamic properties of key reactions involved G protein-coupled receptor (GPCR) signaling from the initial ligand binding step to the generation of the second messenger, cAMP. Well suited to determine rate-limiting reactions taking place along a GPCR signaling cascade in live cells, these techniques have also uncovered new concepts in GPCR signaling as well as many interesting mechanistic subtleties by which GPCRs transmit neurotransmitter and hormone signals into cells.
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2Cyclic AMP - metabolism
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17Membrane proteins
18Microscopy, Confocal
19Microscopy, Fluorescence
20Parathyroid Hormone - metabolism
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17Membrane proteins
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20Parathyroid Hormone - metabolism
21Parathyroid hormone receptor
22Photobleaching
23Protein Binding
24Protein Stability
25Receptors, Parathyroid Hormone - genetics
26Receptors, Parathyroid Hormone - metabolism
27Signal transduction
28Signal Transduction - genetics
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abstractWe describe optical and microscopy methods based on Förster resonance energy transfer, fluorescence recovery after photobleaching, and imaging cross-correlation spectroscopy that permit to determine kinetic and dynamic properties of key reactions involved G protein-coupled receptor (GPCR) signaling from the initial ligand binding step to the generation of the second messenger, cAMP. Well suited to determine rate-limiting reactions taking place along a GPCR signaling cascade in live cells, these techniques have also uncovered new concepts in GPCR signaling as well as many interesting mechanistic subtleties by which GPCRs transmit neurotransmitter and hormone signals into cells.
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pmid23374192
doi10.1016/B978-0-12-407865-9.00016-9
tpages27