Autophagy protein LC3C binding to phospholipid and interaction with lipid membranes.

Uxue Ballesteros, Asier Etxaniz, Marina N Iriondo, Yaiza R Varela, Melisa Lázaro, Ana R Viguera, L Ruth Montes, Mikel Valle, Félix M Goñi, Alicia Alonso

International journal of biological macromolecules 2022 Jul 01

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Autophagy is a process in which parts of the eukaryotic cell are selectively degraded in the lysosome. The materials to be catabolized are first surrounded by a double-membrane structure, the autophagosome. Autophagosome generation is a complex event, in which many proteins are involved. Among the latter, yeast Atg8 or its mammalian orthologues are essential in autophagosome membrane elongation, shaping and closure. A subfamily of the human Atg8 orthologues is formed by the proteins LC3A, LC3B, and LC3C. Previous studies suggest that, at variance with the other two, LC3C does not participate in cardiolipin-mediated mitophagy. The present study was devoted to exploring the binding of LC3C to lipid vesicles, bilayers and monolayers, and the ensuing protein-dependent perturbing effects, in the absence of the mitochondrial lipid cardiolipin. All Atg8 orthologues are covalently bound to a phospholipid prior to their involvement in autophagosome elongation. In our case, a mutant in the C-terminal amino acid, LC3C G126C, together with the use of a maleimide-derivatized phosphatidyl ethanolamine, ensured LC3C lipidation, up to 100% under certain conditions. Ultracentrifugation, surface pressure measurements, spectroscopic and cryo-electron microscopic techniques revealed that lipidated LC3C induced vesicle aggregation (5-fold faster in sonicated than in large unilamellar vesicles) and inter-vesicular lipid mixing (up to 82%), including inner-monolayer lipid mixing (up to 32%), consistent with in vitro partial vesicle fusion. LC3C was also able to cause the release of 80-90% vesicular aqueous contents. The data support the idea that LC3C would be able to help in autophagosome elongation/fusion in autophagy phenomena. Copyright © 2022. Published by Elsevier B.V.

Citation

Uxue Ballesteros, Asier Etxaniz, Marina N Iriondo, Yaiza R Varela, Melisa Lázaro, Ana R Viguera, L Ruth Montes, Mikel Valle, Félix M Goñi, Alicia Alonso. Autophagy protein LC3C binding to phospholipid and interaction with lipid membranes. International journal of biological macromolecules. 2022 Jul 01;212:432-441