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Synthesis of Autophagosomal Marker Protein LC3-II under Detergent-Free Conditions

Authors

  • Yi-Chao Huang,

    1. Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084 (China)
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    • These authors contributed equally to this work.

  • Prof. Dr. Yi-Ming Li,

    1. School of Medical Engineering, Hefei University of Technology, Hefei, Anhui 230009 (China)
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    • These authors contributed equally to this work.

  • Dr. Yang Chen,

    1. School of Life Sciences, Tsinghua University, Beijing 100084 (China)
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  • Man Pan,

    1. Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084 (China)
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  • Yi-Tong Li,

    1. Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084 (China)
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  • Prof. Dr. Li Yu,

    1. School of Life Sciences, Tsinghua University, Beijing 100084 (China)
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  • Prof. Qing-Xiang Guo,

    1. Department of Chemistry, University of Science and Technology of China, Hefei 230026 (China)
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  • Prof. Dr. Lei Liu

    Corresponding author
    1. Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084 (China)
    • Tsinghua-Peking Center for Life Sciences, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084 (China)
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  • This work was supported by the National Basic Research Program of China (973 program No. 2013CB932800) and NSFC (No. 21225207, 20932006, 21102083).

Abstract

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Just add oil: A new detergent-free method was developed to synthesize lipidated proteins using a light-activatable solubilizing side chain (in dashed circle, see scheme) to assist the ligation of the lipopeptides. This method allows the efficient preparation of a phosphatidylethanolamine-conjugated autophagosomal marker protein (LC3-II) as well as labeled derivatives of LC3-II, which can be used to study autophagy regulation.

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