bioRxiv preprint doi: https://doi.org/10.1101/2021.02.07.430153; this version posted February 10, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Cardiac RyR N-terminal region biosensors for FRET-based high-throughput screening Jingyan Zhang1, Siobhan M. Wong King Yuen,2 Jacob A. Schwarz1, Levy M. Treinen1, Ching-Chieh Tung2, Robyn T. Rebbeck1, Kaja Berg3, Bengt Svensson1, Courtney C. Aldrich3, David D. Thomas1, Filip Van Petegem2, and Razvan L. Cornea1* 1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA. 2Department of Biochemistry and Molecular Biology, Life Sciences Institute, University of British Columbia,V6T 1Z3 Vancouver, BC, Canada. 3Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN, 55455, USA. Corresponding author: Razvan L. Cornea Email:
[email protected] Running title: FRET biosensor constructs for RyR-targeted drug discovery Keywords: Ryanodine receptor; N-terminal region; FRET; fluorescence lifetime; high-throughput screening; myopathy. bioRxiv preprint doi: https://doi.org/10.1101/2021.02.07.430153; this version posted February 10, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract failure (HF) (9-13). Physiologically, the opening and closing of RyR channels are tightly regulated The N-terminal region (NTR) of the ryanodine by small molecules, ions, and proteins(14), many receptor (RyR) calcium channels is critical to the of them binding to the enormous RyR cytoplasmic 2+ regulation of Ca release during excitation- portion, with functional effects allosterically contraction coupling.