Abstract
Acetylation of Atg3 regulates the lipidation of the protein Atg8 in autophagy. The molecular mechanism behind this important biochemical event remains to be elucidated. We describe the first semi-synthesis of homogeneous K19/K48-diacetylated Atg3 through sequential hydrazide-based native chemical ligation. In vitro reconstitution experiments with the semi-synthetic proteins confirm that Atg3 acetylation can promote the lipidation of Atg8. We find that acetylation of Atg3 enhances its binding to phosphatidylethanolamine-containing liposomes and to endoplasmic reticulum, through which it promotes the lipidation process.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acetylation
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Amino Acid Sequence
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Autophagy-Related Protein 8 Family / metabolism*
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Autophagy-Related Proteins / chemistry
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Autophagy-Related Proteins / metabolism*
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Endoplasmic Reticulum / metabolism
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Esters / metabolism
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Intracellular Membranes / metabolism*
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Lipids / chemistry*
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Liposomes
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Lysine / metabolism
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Models, Biological
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Protein Binding
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / metabolism*
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Ubiquitin-Conjugating Enzymes / chemistry
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Ubiquitin-Conjugating Enzymes / metabolism*
Substances
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ATG8 protein, S cerevisiae
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Autophagy-Related Protein 8 Family
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Autophagy-Related Proteins
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Esters
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Lipids
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Liposomes
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Saccharomyces cerevisiae Proteins
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Ubiquitin-Conjugating Enzymes
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ATG3 protein, S cerevisiae
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Lysine