Probing the Effect of Sildenafil on Progesterone and Testosterone Production by an Intracellular FRET/BRET Combined Approach.
Animals
Arsenicals
/ chemistry
Biosensing Techniques
/ methods
Catalytic Domain
Cell Line, Tumor
Chorionic Gonadotropin
/ pharmacology
Cyclic AMP
/ metabolism
Cyclic GMP
/ metabolism
Cyclic Nucleotide Phosphodiesterases, Type 5
/ chemistry
Cysteine
/ chemistry
Fluorescence Resonance Energy Transfer
/ methods
Fluorescent Dyes
/ chemistry
HEK293 Cells
Humans
Luteinizing Hormone
/ pharmacology
Mice
Phosphodiesterase 5 Inhibitors
/ metabolism
Progesterone
/ biosynthesis
Protein Binding
Rhodamines
/ chemistry
Sildenafil Citrate
/ metabolism
Testosterone
/ biosynthesis
Journal
Biochemistry
ISSN: 1520-4995
Titre abrégé: Biochemistry
Pays: United States
ID NLM: 0370623
Informations de publication
Date de publication:
12 02 2019
12 02 2019
Historique:
pubmed:
12
12
2018
medline:
1
10
2019
entrez:
12
12
2018
Statut:
ppublish
Résumé
Forster resonance energy transfer (FRET)-based biosensors have been recently applied to the study of biological pathways. In this study, a new biosensor was validated for the first time in live HEK293 and steroidogenic MLTC-1 cell lines for studying the effect of the PDE5 inhibitor on the hCG/LH-induced steroidogenic pathway. The sensor improves FRET between a donor (D), the fluorescein-like diarsenical probe that can covalently bind a tetracysteine motif fused to the PDE5 catalytic domain, and an acceptor (A), the rhodamine probe conjugated to the pseudosubstrate cGMPS. Affinity constant ( K
Identifiants
pubmed: 30532959
doi: 10.1021/acs.biochem.8b01073
doi:
Substances chimiques
Arsenicals
0
Chorionic Gonadotropin
0
Fluorescent Dyes
0
Phosphodiesterase 5 Inhibitors
0
Rhodamines
0
Testosterone
3XMK78S47O
Progesterone
4G7DS2Q64Y
Luteinizing Hormone
9002-67-9
Sildenafil Citrate
BW9B0ZE037
Cyclic AMP
E0399OZS9N
Cyclic Nucleotide Phosphodiesterases, Type 5
EC 3.1.4.35
Cyclic GMP
H2D2X058MU
Cysteine
K848JZ4886
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM