FIGURE 1:

Tepsin directly and specifically binds LC3B in vitro. (A) Schematic diagrams of tepsin and AP-4 depicting the structural basis for the AP-4/tepsin interaction (PDB: 2MJ7); the predicted LIR motif (iLIR database; Jacomin et al., 2016) lies in the unstructured region between the tepsin ENTH (PDB: 5WF9) and VHS-like domains (PDB: 5WF2). (B) Coomassie-stained SDS–PAGE gel and Western blot (α-His; Abcam ab184607) of GST pulldowns using recombinant full-length tepsin-GST (residues 1-525) with His6x-LC3B, His6x-GABARAP (GRP), or His6x-GABARAPL2 (GPL2). Experiments show tepsin binds LC3B and weakly binds GABARAP; free GST was used as a negative control. Representative of three independent experiments.