title: Identification of a Novel Protein Kinase A Inhibitor by Bioluminescence-Based Screening.
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.
authors.affiliation: Department of Molecular Neuroscience, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Ishimoto
authors.forename: Kenji
authors.initials: K
authors.lastname: Azechi
authors.forename: Hisashi
authors.initials: H
authors.lastname: Mori
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism*; D020134:Catalytic Domain; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis / Q000494:pharmacology*; D010766:Phosphorylation; D054730:Protein Interaction Domains and Motifs; D047428:Protein Kinase Inhibitors / Q000032:analysis / Q000494:pharmacology*; D011720:Pyrazoles / Q000032:analysis / Q000494:pharmacology*; D013449:Sulfonamides / Q000032:analysis / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D047908:Intracellular Signaling Peptides and Proteins; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D013449:Sulfonamides; C015482:protein kinase modulator; D005576:Colforsin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D000262:Adenylyl Cyclases