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Taking advantage of the recruitment of Parkin to mitochondria, we next sought to identify the E2 enzymes interacting with Parkin on damaged mitochondria. For this purpose, we utilized fluorescence-based technology detecting PPIs (Fluoppi), which uses multivalent interactions to detect PPIs as fluorescent foci in cells ( 13 Koyano F. Okatsu K. Kosako H. Tamura Y. Go E. Kimura M. Kimura Y. Tsuchiya H. Yoshihara H. Hirokawa T. Endo T. Fon E.A. Trempe J.F. Saeki Y. Tanaka K. Matsuda N.
Mitochondria with decreased membrane potential are characterized by defects in protein import into the matrix and impairments in high-efficiency synthesis of ATP. These low-quality mitochondria are marked with ubiquitin for selective degradation. Key factors in this mechanism are PTEN-induced putative kinase 1 (PINK1, a mitochondrial kinase) and Parkin (a ubiquitin ligase), disruption of which has been implicated in predisposition to Parkinson’s disease.
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