Oxidation of protein kinase a regulatory subunit PKARIα protects against myocardial ischemia-reperfusion injury by inhibiting lysosomal-triggered calcium release
| Autor: | Gerard A Marchal, Keith M. Channon, Oliver Lomas, Raja Jayaram, Besarte Vrellaku, Parag R Gajendragadkar, Nadiia Rawlings, Barbara Casadei, Pawel Swietach, Jillian N. Simon, Larissa Fabritz, Philip Eaton, Dominic Waithe, Sandy Chu, Rana Sayeed, Stefania Monterisi, Fahima Syeda, Manuela Zaccolo |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Cyclic AMP-Dependent Protein Kinase RIalpha Subunit
Protein subunit chemistry.chemical_element Myocardial Reperfusion Injury 030204 cardiovascular system & hematology Calcium calcium signaling protein kinase A phosphorylation Mice 03 medical and health sciences 0302 clinical medicine Original Research Articles Physiology (medical) Lysosome medicine Animals Humans Protein kinase A 030304 developmental biology Calcium signaling 0303 health sciences Kinase business.industry Ryanodine Receptor Calcium Release Channel reperfusion injury medicine.disease Cyclic AMP-Dependent Protein Kinases Cell biology medicine.anatomical_structure chemistry redox ComputingMethodologies_DOCUMENTANDTEXTPROCESSING lysosome Lysosomes Cardiology and Cardiovascular Medicine business Oxidation-Reduction Reperfusion injury Intracellular |
| Zdroj: | Circulation |
| Popis: | Supplemental Digital Content is available in the text. Background: Kinase oxidation is a critical signaling mechanism through which changes in the intracellular redox state alter cardiac function. In the myocardium, PKARIα (type-1 protein kinase A) can be reversibly oxidized, forming interprotein disulfide bonds in the holoenzyme complex. However, the effect of PKARIα disulfide formation on downstream signaling in the heart, particularly under states of oxidative stress such as ischemia and reperfusion (I/R), remains unexplored. Methods: Atrial tissue obtained from patients before and after cardiopulmonary bypass and reperfusion and left ventricular (LV) tissue from mice subjected to I/R or sham surgery were used to assess PKARIα disulfide formation by immunoblot. To determine the effect of disulfide formation on PKARIα catalytic activity and subcellular localization, live-cell fluorescence imaging and stimulated emission depletion super-resolution microscopy were performed in prkar1 knock-out mouse embryonic fibroblasts, neonatal myocytes, or adult LV myocytes isolated from “redox dead” (Cys17Ser) PKARIα knock-in mice and their wild-type littermates. Comparison of intracellular calcium dynamics between genotypes was assessed in fura2-loaded LV myocytes, whereas I/R-injury was assessed ex vivo. Results: In both humans and mice, myocardial PKARIα disulfide formation was found to be significantly increased (2-fold in humans, P=0.023; 2.4-fold in mice, P |
| Databáze: | OpenAIRE |
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