An ultrasensitive biosensor for high-resolution kinase activity imaging in awake mice

Jin Fan Zhang, Bian Liu, Ingie Hong, Albert Mo, Richard H. Roth, Brian Tenner, Wei Lin, Jason Z. Zhang, Rosana S. Molina, Mikhail Drobizhev, Thomas E. Hughes, Lin Tian, Richard L. Huganir, Sohum Mehta, Jin Zhang

Research output: Contribution to journalArticlepeer-review

Abstract

Protein kinases control nearly every facet of cellular function. These key signaling nodes integrate diverse pathway inputs to regulate complex physiological processes, and aberrant kinase signaling is linked to numerous pathologies. While fluorescent protein-based biosensors have revolutionized the study of kinase signaling by allowing direct, spatiotemporally precise kinase activity measurements in living cells, powerful new molecular tools capable of robustly tracking kinase activity dynamics across diverse experimental contexts are needed to fully dissect the role of kinase signaling in physiology and disease. Here, we report the development of an ultrasensitive, second-generation excitation-ratiometric protein kinase A (PKA) activity reporter (ExRai-AKAR2), obtained via high-throughput linker library screening, that enables sensitive and rapid monitoring of live-cell PKA activity across multiple fluorescence detection modalities, including plate reading, cell sorting and one- or two-photon imaging. Notably, in vivo visual cortex imaging in awake mice reveals highly dynamic neuronal PKA activity rapidly recruited by forced locomotion. [Figure not available: see fulltext.]

Original languageEnglish (US)
Pages (from-to)39-46
Number of pages8
JournalNature chemical biology
Volume17
Issue number1
DOIs
StatePublished - Jan 2021

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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