Fig. 7: Factors affecting Las17 interactions.
A Microscale thermophoresis showing binding of Las17 to actin in the presence (red) or absence (blue) of Sla1 SH3 domains #1–3. The concentration of actin (50 nM) and Sla1 (4.5 µM) were kept constant throughout the experiment, and the concentration of Las17 300–422 was varied between 0.29 and 9.5 µM. Error bars are standard deviation. B Liposome co-sedimentation assay shows that GST-Sla1 SH3#1-3 co-precipitates with liposomes prepared from bovine brain extract, whereas GST alone does not. C Quantification of liposome co-sedimentation assays. Å Ãdák’s multiple comparisons test P < 0.0001 (n = 6). D Representative pyrene actin assay showing alleviation of Sla1 inhibition (red) by Sec4 (pink). Actin only (black); Actin + Las17 (blue). E Alphafold prediction of an interaction of Sec4 with a region of Las17 primarily between 327 and 333 which lies at the C-terminal end of the first Las17 PP motif. Shown is surface representation of Sec4 in green and ribbon depiction of Las17 in blue with predicted interacting surface residues within 3.5 Å in red.
