Chen Qun, Li Ling, Samidurai Arun, Thompson Jeremy, Hu Ying, Willard Belinda, Lesnefsky Edward J
Department of Internal Medicine, Division of Cardiology, Pauley Heart Center, Virginia Commonwealth University, Richmond, Virginia, USA.
Proteomics Core, Cleveland Clinic, Cleveland, Ohio, USA.
FASEB J. 2024 Jan 31;38(2):e23404. doi: 10.1096/fj.202301158RR.
The induction of acute endoplasmic reticulum (ER) stress damages the electron transport chain (ETC) in cardiac mitochondria. Activation of mitochondria-localized calpain 1 (CPN1) and calpain 2 (CPN2) impairs the ETC in pathological conditions, including aging and ischemia-reperfusion in settings where ER stress is increased. We asked if the activation of calpains causes the damage to the ETC during ER stress. Control littermate and CPNS1 (calpain small regulatory subunit 1) deletion mice were used in the current study. CPNS1 is an essential subunit required to maintain CPN1 and CPN2 activities, and deletion of CPNS1 prevents their activation. Tunicamycin (TUNI, 0.4 mg/kg) was used to induce ER stress in C57BL/6 mice. Cardiac mitochondria were isolated after 72 h of TUNI treatment. ER stress was increased in both control littermate and CPNS1 deletion mice with TUNI treatment. The TUNI treatment activated both cytosolic and mitochondrial CPN1 and 2 (CPN1/2) in control but not in CPNS1 deletion mice. TUNI treatment led to decreased oxidative phosphorylation and complex I activity in control but not in CPNS1 deletion mice compared to vehicle. The contents of complex I subunits, including NDUFV2 and ND5, were decreased in control but not in CPNS1 deletion mice. TUNI treatment also led to decreased oxidation through cytochrome oxidase (COX) only in control mice. Proteomic study showed that subunit 2 of COX was decreased in control but not in CPNS1 deletion mice. Our results provide a direct link between activation of CPN1/2 and complex I and COX damage during acute ER stress.
急性内质网(ER)应激的诱导会损害心脏线粒体中的电子传递链(ETC)。线粒体定位的钙蛋白酶1(CPN1)和钙蛋白酶2(CPN2)的激活会在包括衰老和内质网应激增加情况下的缺血再灌注等病理条件下损害ETC。我们询问钙蛋白酶的激活是否会在内质网应激期间导致ETC损伤。本研究使用了对照同窝小鼠和CPNS1(钙蛋白酶小调节亚基1)缺失小鼠。CPNS1是维持CPN1和CPN2活性所需的必需亚基,CPNS1的缺失可阻止它们的激活。衣霉素(TUNI,0.4mg/kg)用于诱导C57BL/6小鼠的内质网应激。TUNI处理72小时后分离心脏线粒体。TUNI处理后,对照同窝小鼠和CPNS1缺失小鼠的内质网应激均增加。TUNI处理在对照小鼠中激活了胞质和线粒体中的CPN1和2(CPN1/2),但在CPNS1缺失小鼠中未激活。与溶媒相比,TUNI处理导致对照小鼠而非CPNS1缺失小鼠的氧化磷酸化和复合体I活性降低。对照小鼠中复合体I亚基(包括NDUFV2和ND5)的含量降低,但CPNS1缺失小鼠中未降低。TUNI处理还仅在对照小鼠中导致细胞色素氧化酶(COX)的氧化减少。蛋白质组学研究表明,对照小鼠中COX的亚基2减少,但CPNS1缺失小鼠中未减少。我们的结果提供了急性内质网应激期间CPN1/2激活与复合体I和COX损伤之间的直接联系。