The PTEN-induced kinase 1 (PINK1)-Parkin pathway plays a vital role in maintaining a healthy pool of mitochondria in higher eukaryotic cells. While the downstream components of this pathway are well understood, the upstream triggers remain less explored. In this study, we conducted an extensive analysis of inhibitors targeting various mitochondrial electron transport chain (ETC) complexes to investigate their potential as activators of the PINK1–Parkin pathway. We identified cloflucarban, an antibacterial compound, as a novel pathway activator that simultaneously inhibits mitochondrial complexes III and V, and V. RNA interference (RNAi) confirmed that the dual inhibition of these complexes activates the PINK1–Parkin pathway. Intriguingly, we discovered that albumin, specifically bovine serum albumin (BSA) and human serum albumin (HSA) commonly present in culture media, can hinder carbonyl cyanide m-chlorophenyl hydrazone (CCCP)-induced pathway activation. However, cloflucarban’s efficacy remains unaffected by albumin, highlighting its reliability for studying the PINK1–Parkin pathway. This study provides insights into the activation of the upstream PINK1–Parkin pathway and underscores the influence of culture conditions on research outcomes. Cloflucarban emerges as a promising tool for investigating mitochondrial quality control and neurodegenerative diseases.

中文翻译：

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氯氟卡班通过线粒体复合物抑制阐明 PINK1-Parkin 通路的特异性和环境依赖性激活

PTEN 诱导的激酶 1 (PINK1)-Parkin 通路在维持高等真核细胞中线粒体的健康池中发挥着至关重要的作用。虽然该途径的下游组成部分已被充分了解，但上游触发因素的研究仍较少。在这项研究中，我们对针对各种线粒体电子传递链 (ETC) 复合物的抑制剂进行了广泛的分析，以研究它们作为 PINK1-Parkin 通路激活剂的潜力。我们确定了氯氟卡班（一种抗菌化合物）作为一种新型通路激活剂，可同时抑制线粒体复合物 III、V 和 V。RNA 干扰 (RNAi) 证实，这些复合物的双重抑制可激活 PINK1-Parkin 通路。有趣的是，我们发现白蛋白，特别是培养基中常见的牛血清白蛋白（BSA）和人血清白蛋白（HSA），可以阻碍羰基氰化物间氯苯腙（CCCP）诱导的途径激活。然而，氯氟卡班的功效仍然不受白蛋白的影响，突显了其在研究 PINK1-Parkin 通路方面的可靠性。这项研究提供了对上游 PINK1-Parkin 通路激活的见解，并强调了培养条件对研究结果的影响。氯氟卡班成为研究线粒体质量控制和神经退行性疾病的有前途的工具。