Fig. 7: Tim44 C-terminus is involved in presequence interaction.
From: Mapping protein interactions in the active TOM-TIM23 supercomplex
a Isolated mitochondria were incubated with presequence photo-peptides pL19B and pL16B in the presence or absence of membrane potential (ΔΨ) and subjected to UV irradiation. Samples were analyzed by immunoblotting. PA: photo-adduct. Three independent replicates were carried out. Recombinantly purified b full length Tim44 and c Tim44C-term were subjected to photo-peptide incubation and UV irradiation. Analysis of photo-adduct formation was carried out by SDS-PAGE and colloidal coomassie staining or immunoblotting. SA-HRP: HRP conjugated Streptavidin. Representative images from two independent experiments. Purified His6-tagged d Tim44 full length and e Tim44C-term were analyzed by SPR (Surface Plasma Resonance) analysis at increasing analyte concentrations (0.125−16 µM). A representative sensogram is depicted. Black lines show observed binding, orange calculated ka/kd fitting of the kinetic data. Analytes used: presequence peptide pALDH or scrambled presequence pALDH-s.
