한빛사논문
- 조회669
Jin-Fan Zhang1,2,9, Bian Liu 3,4,9, Ingie Hong 3,4,9, Albert Mo1, Richard H. Roth 3,4,8, Brian Tenner1, Wei Lin1, Jason Z. Zhang 1,2, Rosana S. Molina 5, Mikhail Drobizhev5, Thomas E. Hughes5, Lin Tian 6, Richard L. Huganir 3,4, Sohum Mehta 1 and Jin Zhang 1,2,7
1Department of Pharmacology, University of California, San Diego, La Jolla, CA, USA.
2Department of Bioengineering, University of California, San Diego, La Jolla, CA, USA.
3The Solomon H. Snyder Department of Neuroscience, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
4The Kavli Neuroscience Discovery Institute, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
5Department of Cell Biology and Neuroscience, Montana State University, Bozeman, MT, USA.
6Department of Biochemistry and Molecular Medicine, University of California, Davis, Sacramento, CA, USA.
7Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
8Present address: Department of Neurosurgery, Stanford University School of Medicine, Palo Alto, CA, USA.
9These authors contributed equally: Jin-Fan Zhang, Bian Liu, Ingie Hong.
Corresponding authors : Correspondence to Richard L. Huganir, Sohum Mehta or Jin Zhang.
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.
논문정보
- Research article
- 2021년 01월 (BRIC 등록일 2023-07-14)
- 국외 연구진
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