Rapid subcellular calcium responses and dynamics by calcium sensor G-CatchER+
Florence N. Reddish,
Cassandra L. Miller,
Xiaonan Deng,
Bin Dong,
Atit A. Patel,
Mohammad A. Ghane,
Barbara Mosca,
Cheyenne McBean,
Shengnan Wu,
Kyril M. Solntsev,
You Zhuo,
Giovanni Gadda,
Ning Fang,
Daniel N. Cox,
Angela M. Mabb,
Susan Treves,
Francesco Zorzato,
Jenny J. Yang
Affiliations
Florence N. Reddish
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Cassandra L. Miller
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Xiaonan Deng
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Bin Dong
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Atit A. Patel
Neuroscience Institute, Georgia State University, Atlanta, GA 30303, USA; Center for Behavioral Neuroscience, Georgia State University, Atlanta, GA 30303, USA
Mohammad A. Ghane
Neuroscience Institute, Georgia State University, Atlanta, GA 30303, USA; Center for Behavioral Neuroscience, Georgia State University, Atlanta, GA 30303, USA
Barbara Mosca
Department of Life Sciences, General Pathology, University of Ferrara, Ferrara, Italy
Cheyenne McBean
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Shengnan Wu
Center for Molecular and Translational Medicine, Georgia State University, Atlanta, GA 30303, USA
Kyril M. Solntsev
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA
You Zhuo
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Giovanni Gadda
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Ning Fang
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA
Daniel N. Cox
Neuroscience Institute, Georgia State University, Atlanta, GA 30303, USA; Center for Behavioral Neuroscience, Georgia State University, Atlanta, GA 30303, USA
Angela M. Mabb
Neuroscience Institute, Georgia State University, Atlanta, GA 30303, USA; Center for Behavioral Neuroscience, Georgia State University, Atlanta, GA 30303, USA
Susan Treves
Department of Life Sciences, General Pathology, University of Ferrara, Ferrara, Italy; Department of Biomedicine, Basel University, Hebelstrasse 20, 4031 Basel, Switzerland
Francesco Zorzato
Department of Life Sciences, General Pathology, University of Ferrara, Ferrara, Italy; Department of Biomedicine, Basel University, Hebelstrasse 20, 4031 Basel, Switzerland
Jenny J. Yang
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA; Corresponding author
Summary: The precise spatiotemporal characteristics of subcellular calcium (Ca2+) transients are critical for the physiological processes. Here we report a green Ca2+ sensor called “G-CatchER+” using a protein design to report rapid local ER Ca2+ dynamics with significantly improved folding properties. G-CatchER+ exhibits a superior Ca2+ on rate to G-CEPIA1er and has a Ca2+-induced fluorescence lifetimes increase. G-CatchER+ also reports agonist/antagonist triggered Ca2+ dynamics in several cell types including primary neurons that are orchestrated by IP3Rs, RyRs, and SERCAs with an ability to differentiate expression. Upon localization to the lumen of the RyR channel (G-CatchER+-JP45), we report a rapid local Ca2+ release that is likely due to calsequestrin. Transgenic expression of G-CatchER+ in Drosophila muscle demonstrates its utility as an in vivo reporter of stimulus-evoked SR local Ca2+ dynamics. G-CatchER+ will be an invaluable tool to examine local ER/SR Ca2+ dynamics and facilitate drug development associated with ER dysfunction.
