Cell-Free Reconstitution of Autophagic Membrane Formation
Autophagy is a catabolic pathway for bulk turnover of cytoplasmic components through the lysosome. Completion of autophagy requires a sophisticated membrane remodeling process. The early steps involve autophagic membrane precursor generation from the intracellular membranes. The intricate protein-membrane interactions underlying autophagic membrane precursor generation have been a focus of attention but yet poorly defined. Here, we summarize the procedure of a cell-free system we have established to dissect the molecular mechanism of early autophagic membrane generation.
Key wordsAutophagy Autophagosome Cell-free reconstitution LC3 lipidation
L.G. was previously supported by the NIH Pathway to Independence Award (K99/R00) National Institute of General Medical Sciences (Grant Number: 1K99GM114397-02) and is now supported by the State Key Laboratory of Membrane Biology, China; the National Natural Science Foundation of China (31741082), the Tsinghua-Peking Center for Life Sciences, and the National Young Thousand Talents Program. L.G. is deeply grateful for the training provided by Dr. Randy Schekman at the University of California, Berkeley. We thank Bob Lesch at University of California Berkeley for critical reading and helpful editing of the manuscript.
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