Measuring CREB Activation Using Bioluminescent Probes That Detect KID鈥揔IX Interaction in Living Cells

详细信息    查看全文

• 作者：Tetsuya Ishimoto ; Hiroki Mano ; Takeaki Ozawa ; Hisashi Mori
• 刊名：Bioconjugate Chemistry
• 出版年：2012
• 出版时间：May 16, 2012
• 年：2012
• 卷：23
• 期：5
• 页码：923-932
• 全文大小：534K
• 年卷期：v.23,no.5(May 16, 2012)
• ISSN：1520-4812

文摘

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鈥揔IX 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.