Disrupting the PLP – Kinesin-1 interaction. (A) Yeast clones showing the PLP interactions disrupted by the PLP^PD mutation. Note that Asl interaction is also disrupted. Interaction was determined by the growth of blue clones on the selection plate (QDOXA). (B) Yeast clones showing the interactions disrupted by deletion of PLP aa741-970. The interaction with KHC^850-975 appears weaker due to decreased growth. Note interaction with PLP^2539-2895 is also disrupted. (C) Validation of PLP–KHC interaction mutants by mitochondrial targeting assay. PLP fragments were targeted to the mitochondria using the Tom20 mitochondrial localization sequence (red). The cargo binding tail of KHC (KHC^850-975) was tagged with mNeon. Interaction was determined by recruitment of mNeon::KHC^850-975 to the mitochondria. (Arrowheads point to mitochondria; magenta dashed line labels the nuclei.) (D and E) Histograms of single particle mass values determined by mass photometry for HALO::PLP^584-1811: WT and PD mutant, respectively. Lines are the Gaussian fit to the data yielding the molecular weights, consistent with a predominant dimer (346 kD) and a minor tetramer (692 kD) species of PLP. Scale bars: 5 µm.