Unique amphipathic α helix drives membrane insertion and enzymatic activity of ATG3.

Nishimura, Taki; Lazzeri, Gianmarco; Mizushima, Noboru; Covino, Roberto; Tooze, Sharon A · Sci Adv · 2023

basic_science · Level V

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Abstract

Autophagosome biogenesis requires a localized perturbation of lipid membrane dynamics and a unique protein-lipid conjugate. Autophagy-related (ATG) proteins catalyze this biogenesis on cellular membranes, but the underlying molecular mechanism remains unclear. Focusing on the final step of the protein-lipid conjugation reaction, the ATG8/LC3 lipidation, we show how the membrane association of the conjugation machinery is organized and fine-tuned at the atomistic level. Amphipathic α helices in ATG3 proteins (AH<sub>ATG3</sub>) have low hydrophobicity and contain less bulky residues. Molecular dynamics simulations reveal that AH<sub>ATG3</sub> regulates the dynamics and accessibility of the thioester bond of the ATG3~LC3 conjugate to lipids, enabling the covalent lipidation of LC3. Live-cell imaging shows that the transient membrane association of ATG3 with autophagic membranes is governed by the less bulky-hydrophobic feature of AH<sub>ATG3</sub>. The unique properties of AH<sub>ATG3</sub> facilitate protein-lipid bilayer association, leading to the remodeling of the lipid bilayer required for the formation of autophagosomes.

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