Calpain cleavage of Junctophilin-2 generates a spectrum of calcium-dependent cleavage products and DNA-rich NT 1 -fragment domains in cardiomyocytes.

Autor: Weninger G; Cellular Biophysics and Translational Cardiology Section, Heart Research Center Göttingen, University Medical Center Göttingen, Robert-Koch-Str. 42a, 37075, Göttingen, Germany.; Department of Cardiology and Pneumology, University Medical Center Göttingen, 37075, Göttingen, Germany.; Collaborative Research Center SFB1190 'Compartmental Gates and Contact Sites in Cells', University of Göttingen, 37073, Göttingen, Germany.; Department of Physiology and Cellular Biophysics, Center for Molecular Cardiology, Columbia University Vagelos College of Physicians and Surgeons, New York, NY, 10032, USA., Pochechueva T; Cellular Biophysics and Translational Cardiology Section, Heart Research Center Göttingen, University Medical Center Göttingen, Robert-Koch-Str. 42a, 37075, Göttingen, Germany.; Department of Cardiology and Pneumology, University Medical Center Göttingen, 37075, Göttingen, Germany.; Collaborative Research Center SFB1190 'Compartmental Gates and Contact Sites in Cells', University of Göttingen, 37073, Göttingen, Germany., El Chami D; Cellular Biophysics and Translational Cardiology Section, Heart Research Center Göttingen, University Medical Center Göttingen, Robert-Koch-Str. 42a, 37075, Göttingen, Germany.; Department of Cardiology and Pneumology, University Medical Center Göttingen, 37075, Göttingen, Germany.; Collaborative Research Center SFB1190 'Compartmental Gates and Contact Sites in Cells', University of Göttingen, 37073, Göttingen, Germany., Luo X; Institute of Pharmacology and Toxicology, Technische Universität Dresden, 01307, Dresden, Germany., Kohl T; Cellular Biophysics and Translational Cardiology Section, Heart Research Center Göttingen, University Medical Center Göttingen, Robert-Koch-Str. 42a, 37075, Göttingen, Germany.; Department of Cardiology and Pneumology, University Medical Center Göttingen, 37075, Göttingen, Germany.; Collaborative Research Center SFB1190 'Compartmental Gates and Contact Sites in Cells', University of Göttingen, 37073, Göttingen, Germany.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC2067), University of Göttingen, 37073, Göttingen, Germany.; DZHK (German Centre for Cardiovascular Research), partner site, 37075, Göttingen, Germany., Brandenburg S; Cellular Biophysics and Translational Cardiology Section, Heart Research Center Göttingen, University Medical Center Göttingen, Robert-Koch-Str. 42a, 37075, Göttingen, Germany.; Department of Cardiology and Pneumology, University Medical Center Göttingen, 37075, Göttingen, Germany.; Collaborative Research Center SFB1190 'Compartmental Gates and Contact Sites in Cells', University of Göttingen, 37073, Göttingen, Germany.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC2067), University of Göttingen, 37073, Göttingen, Germany.; DZHK (German Centre for Cardiovascular Research), partner site, 37075, Göttingen, Germany., Urlaub H; Collaborative Research Center SFB1190 'Compartmental Gates and Contact Sites in Cells', University of Göttingen, 37073, Göttingen, Germany.; Proteomanalyse, Department of Clinical Chemistry, University Medical Center Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.; Bioanalytical Mass Spectrometry, Max Planck Institute for Multidisciplinary Sciences, 37077, Göttingen, Germany., Guan K; Institute of Pharmacology and Toxicology, Technische Universität Dresden, 01307, Dresden, Germany., Lenz C; Proteomanalyse, Department of Clinical Chemistry, University Medical Center Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany. christof.lenz@med.uni-goettingen.de.; Bioanalytical Mass Spectrometry, Max Planck Institute for Multidisciplinary Sciences, 37077, Göttingen, Germany. christof.lenz@med.uni-goettingen.de., Lehnart SE; Cellular Biophysics and Translational Cardiology Section, Heart Research Center Göttingen, University Medical Center Göttingen, Robert-Koch-Str. 42a, 37075, Göttingen, Germany. slehnart@med.uni-goettingen.de.; Department of Cardiology and Pneumology, University Medical Center Göttingen, 37075, Göttingen, Germany. slehnart@med.uni-goettingen.de.; Collaborative Research Center SFB1190 'Compartmental Gates and Contact Sites in Cells', University of Göttingen, 37073, Göttingen, Germany. slehnart@med.uni-goettingen.de.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC2067), University of Göttingen, 37073, Göttingen, Germany. slehnart@med.uni-goettingen.de.; DZHK (German Centre for Cardiovascular Research), partner site, 37075, Göttingen, Germany. slehnart@med.uni-goettingen.de.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2022 Jun 20; Vol. 12 (1), pp. 10387. Date of Electronic Publication: 2022 Jun 20.
DOI: 10.1038/s41598-022-14320-9
Abstrakt: Calpains are calcium-activated neutral proteases involved in the regulation of key signaling pathways. Junctophilin-2 (JP2) is a Calpain-specific proteolytic target and essential structural protein inside Ca 2+ release units required for excitation-contraction coupling in cardiomyocytes. While downregulation of JP2 by Calpain cleavage in heart failure has been reported, the precise molecular identity of the Calpain cleavage sites and the (patho-)physiological roles of the JP2 proteolytic products remain controversial. We systematically analyzed the JP2 cleavage fragments as function of Calpain-1 versus Calpain-2 proteolytic activities, revealing that both Calpain isoforms preferentially cleave mouse JP2 at R565, but subsequently at three additional secondary Calpain cleavage sites. Moreover, we identified the Calpain-specific primary cleavage products for the first time in human iPSC-derived cardiomyocytes. Knockout of RyR2 in hiPSC-cardiomyocytes destabilized JP2 resulting in an increase of the Calpain-specific cleavage fragments. The primary N-terminal cleavage product NT 1 accumulated in the nucleus of mouse and human cardiomyocytes in a Ca 2+ -dependent manner, closely associated with euchromatic chromosomal regions, where NT 1 is proposed to function as a cardio-protective transcriptional regulator in heart failure. Taken together, our data suggest that stabilizing NT 1 by preventing secondary cleavage events by Calpain and other proteases could be an important therapeutic target for future studies.
(© 2022. The Author(s).)
Databáze: MEDLINE
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