Design and expression of various PINK1 constructs. A) Three endogenous PINK1 forms can be detected and identified in Hela cells using the Odyssey Infrared Imaging. Valinomycin (Δψm dissipater) increases PINK1 FL, epoxomicin (proteasome inhibitor) increases both PINK1 Δ1 and Δ2. A') PINK1 siRNA knocked down all three endogenous PINK1 forms. Asterisk denotes non-specific band from α-PINK1 antibodies. B) Schematic diagram of various PINK1 deletion mutants and various MLS-PINK1 fusion constructs. Cyt b2 is yeast cytochrome b2 MLS from 1-33 amino acids. IMMT is human mitofilin MLS from 1-187 amino acids. C) Protein expression of aforementioned PINK1 constructs relative to WT PINK1 is compared by anti-FLAG western blot. All PINK1 deletion mutants express PINK1 Δ2 form, corresponding to Δ151 PINK1 molecular weight. Immt-Δ151 PINK1 is detected as a single protein product.
Lin and Kang BMC Cell Biology 2010 11:90 doi:10.1186/1471-2121-11-90