Abstract
We screened inhibitors in the adenylyl cyclase/protein kinase A/cAMP response element binding protein pathway (AC/PKA/CREB pathway) from a 2400 chemical library by a cell-based assay method using bioluminescence probes. We found a compound that inhibited forskolin-induced cAMP response element (CRE)-dependent transcription, the interaction between the kinase-inducible domain (KID) and the interacting domain (KIX), and endogenous CREB phosphorylation. Furthermore, this compound suppressed the activity of the PKA catalytic subunit dose-dependently. On the other hand, this compound did not inhibit forskolin-induced cAMP up-regulation. Taken together, we conclude that we have identified a new PKA inhibitor that binds to the catalytic subunit directly. We also succeeded in shortening the screening protocol by excluding a screening step which was used in a previous method.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenylyl Cyclases / metabolism*
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Catalytic Domain
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Colforsin / pharmacology
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Cyclic AMP / metabolism*
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Cyclic AMP Response Element-Binding Protein / metabolism*
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Cyclic AMP-Dependent Protein Kinases / metabolism*
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HEK293 Cells
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Humans
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Intracellular Signaling Peptides and Proteins / analysis
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Intracellular Signaling Peptides and Proteins / pharmacology*
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Phosphorylation
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Protein Interaction Domains and Motifs
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Protein Kinase Inhibitors / analysis
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Protein Kinase Inhibitors / pharmacology*
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Pyrazoles / analysis
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Pyrazoles / pharmacology*
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Sulfonamides / analysis
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Sulfonamides / pharmacology*
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Transcription, Genetic / drug effects
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Up-Regulation
Substances
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Cyclic AMP Response Element-Binding Protein
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Intracellular Signaling Peptides and Proteins
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Protein Kinase Inhibitors
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Pyrazoles
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Sulfonamides
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protein kinase modulator
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Colforsin
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Cyclic AMP
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Cyclic AMP-Dependent Protein Kinases
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Adenylyl Cyclases