Fig. 5: The structural basis of the Atg40–Atg8 interaction.
a Amino-acid sequences of the canonical AIM and the C-terminal region of Atg40 (230–256). The residues colored red were replaced with alanine in c. b Crystal structure of Atg8-Atg40237–252. Yellow dashed lines, salt bridges. c ATG8-GFP atg4Δ atg40Δ cells expressing Atg40-HA were treated with rapamycin for 6 h, and the cell lysates (input) were subjected to immunoprecipitation using anti-GFP antibody. The immunoprecipitates (IP) are analyzed by immunoblotting using antibodies against GFP and HA. Additional bands in Atg40-HA immunoblotting represent its degradation products (deg.). The experiments were repeated independently twice. d Binding affinity of Atg40 peptides to Atg8 was measured by ITC.
