A semisynthetic Atg3 reveals that acetylation promotes Atg3 membrane binding and Atg8 lipidation.
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.
School of Biological and Medical Engineering, Hefei University of Technology, Anhui, Hefei 230009, China.|Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry &Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084, China.
Yi-Tong
YT
Li
Tsinghua-Peking Center for Life Sciences, Key Laboratory of Biomembrane and Membrane Biotechnology, Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Cong
C
Yi
School of Biological and Medical Engineering, Hefei University of Technology, Anhui, Hefei 230009, China.
Chen-Chen
CC
Chen
Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry &Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084, China.
Huan
H
Lan
Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry &Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084, China.
Man
M
Pan
Tsinghua-Peking Center for Life Sciences, Key Laboratory of Biomembrane and Membrane Biotechnology, Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Shao-Jin
SJ
Zhang
Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry &Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084, China.
Yi-Chao
YC
Huang
School of Biological and Medical Engineering, Hefei University of Technology, Anhui, Hefei 230009, China.
Chao-Jian
CJ
Guan
School of Biological and Medical Engineering, Hefei University of Technology, Anhui, Hefei 230009, China.
Yi-Ming
YM
Li
Tsinghua-Peking Center for Life Sciences, Key Laboratory of Biomembrane and Membrane Biotechnology, Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Li
L
Yu
Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry &Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084, China.
Lei
L
Liu
D000107:Acetylation; D000595:Amino Acid Sequence; D000071190:Autophagy-Related Protein 8 Family / Q000378:metabolism*; D000071183:Autophagy-Related Proteins / Q000737:chemistry / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004952:Esters / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism*; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes; D008239:Lysine / Q000378:metabolism; D008954:Models, Biological; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000378:metabolism*
C113757:ATG8 protein, S cerevisiae; D000071190:Autophagy-Related Protein 8 Family; D000071183:Autophagy-Related Proteins; D004952:Esters; D008055:Lipids; D008081:Liposomes; D029701:Saccharomyces cerevisiae Proteins; D044763:Ubiquitin-Conjugating Enzymes; C104235:ATG3 protein, S cerevisiae; D008239:Lysine