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

doi:10.1016/j.isci.2021.102129

Rapid subcellular calcium responses and dynamics by calcium sensor G-CatchER

iScience. 2021 Feb 3;24(3):102129. doi: 10.1016/j.isci.2021.102129. eCollection 2021 Mar 19.

Authors

Florence N Reddish¹, Cassandra L Miller¹, Xiaonan Deng¹, Bin Dong¹, Atit A Patel^{2

3}, Mohammad A Ghane^{2

3}, Barbara Mosca⁴, Cheyenne McBean¹, Shengnan Wu⁵, Kyril M Solntsev⁶, You Zhuo¹, Giovanni Gadda¹, Ning Fang¹, Daniel N Cox^{2

3}, Angela M Mabb^{2

3}, Susan Treves^{4

7}, Francesco Zorzato^{4

7}, Jenny J Yang¹

Affiliations

¹ Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA.
² Neuroscience Institute, Georgia State University, Atlanta, GA 30303, USA.
³ Center for Behavioral Neuroscience, Georgia State University, Atlanta, GA 30303, USA.
⁴ Department of Life Sciences, General Pathology, University of Ferrara, Ferrara, Italy.
⁵ Center for Molecular and Translational Medicine, Georgia State University, Atlanta, GA 30303, USA.
⁶ School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA.
⁷ Department of Biomedicine, Basel University, Hebelstrasse 20, 4031 Basel, Switzerland.

Abstract

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.

Keywords: biochemistry; biophysics; cell biology.

Abstract

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