Development of a Novel SNAP-Epitope Tag/Near-Infrared Imaging Assay to Quantify G Protein-Coupled Receptor Degradation in Human Cells.
Bortezomib
/ pharmacology
Cell Line, Tumor
Cycloheximide
/ pharmacology
Epithelial Cells
/ cytology
Gene Expression
HEK293 Cells
Half-Life
High-Throughput Screening Assays
Humans
Molecular Imaging
/ methods
Norepinephrine
/ pharmacology
Proteasome Endopeptidase Complex
/ drug effects
Proteasome Inhibitors
/ pharmacology
Protein Isoforms
/ genetics
Protein Synthesis Inhibitors
/ pharmacology
Proteolysis
/ drug effects
Receptor, Serotonin, 5-HT2A
/ genetics
Receptors, Adrenergic, alpha-1
/ genetics
Receptors, Interleukin-8A
/ genetics
Receptors, Somatostatin
/ genetics
Recombinant Fusion Proteins
/ chemistry
Spectroscopy, Near-Infrared
/ methods
G protein-coupled receptor
SNAP
cycloheximide-chase
degradation
label-free dynamic mass redistribution
Journal
SLAS discovery : advancing life sciences R & D
ISSN: 2472-5560
Titre abrégé: SLAS Discov
Pays: United States
ID NLM: 101697563
Informations de publication
Date de publication:
04 2021
04 2021
Historique:
pubmed:
7
1
2021
medline:
1
3
2022
entrez:
6
1
2021
Statut:
ppublish
Résumé
We have developed a novel reporter assay that leverages SNAP-epitope tag/near-infrared (NIR) imaging technology to monitor G protein-coupled receptor (GPCR) degradation in human cell lines. N-terminal SNAP-tagged GPCRs were subcloned and expressed in human embryonic kidney (HEK) 293 cells and then subjected to 24 h of cycloheximide (CHX)-chase degradation assays to quantify receptor degradation half-lives (
Identifiants
pubmed: 33402011
doi: 10.1177/2472555220979793
pmc: PMC9246501
mid: NIHMS1816542
pii: S2472-5552(22)06698-9
doi:
Substances chimiques
ADRA1A protein, human
0
Proteasome Inhibitors
0
Protein Isoforms
0
Protein Synthesis Inhibitors
0
Receptor, Serotonin, 5-HT2A
0
Receptors, Adrenergic, alpha-1
0
Receptors, Interleukin-8A
0
Receptors, Somatostatin
0
Recombinant Fusion Proteins
0
somatostatin receptor type 1
0
Bortezomib
69G8BD63PP
Cycloheximide
98600C0908
Proteasome Endopeptidase Complex
EC 3.4.25.1
Norepinephrine
X4W3ENH1CV
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
570-578Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM100893
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007750
Pays : United States
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