title: Measuring CREB activation using bioluminescent probes that detect KID-KIX interaction in living cells.
abstract: The cyclic adenosine monophosphate response element-binding protein (CREB) is a transcription factor that contributes to memory formation. The transcriptional activity of CREB is induced by its phosphorylation at Ser-133 and subsequent interaction with the CREB-binding protein (CBP)/p300. We designed and optimized firefly split luciferase probe proteins that detect the interaction of the kinase-inducible domain (KID) of CREB and the KIX domain of CBP/p300. The increase in the light intensity of the probe proteins results from the phosphorylation of the responsible serine corresponding to Ser-133 of CREB. Because these proteins have a high signal-to-noise ratio and are nontoxic, it has become possible for the first time to carry out long-term measurement of KID-KIX interaction in living cells. Furthermore, we examined the usefulness of the probe proteins for future high-throughput cell-based drug screening and found several herbal extracts that activated CREB.
authors.affiliation: Department of Molecular Neuroscience, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama , Toyama, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Ishimoto
authors.forename: Hiroki
authors.initials: H
authors.lastname: Mano
authors.forename: Takeaki
authors.initials: T
authors.lastname: Ozawa
authors.forename: Hisashi
authors.initials: H
authors.lastname: Mori
mesh_terms: D001665:Binding Sites; D017362:Cyclic AMP Response Element-Binding Protein / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D049409:Luciferases, Firefly / Q000032:analysis / Q000378:metabolism*; D049408:Luminescent Agents / Q000032:analysis / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods; D010766:Phosphorylation; D010936:Plant Extracts / Q000494:pharmacology; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping / Q000379:methods*; D050880:p300-CBP Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D049408:Luminescent Agents; D010936:Plant Extracts; D049409:Luciferases, Firefly; D050880:p300-CBP Transcription Factors