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Ma Xuejing, Zhi Zelun, Zhang Shuyan, Liu Pingsheng. Measurement of ATGL activity using adiposomes[J]. Biophysics Reports. doi: 10.52601/bpr.2023.220016
Citation: Ma Xuejing, Zhi Zelun, Zhang Shuyan, Liu Pingsheng. Measurement of ATGL activity using adiposomes[J]. Biophysics Reports. doi: 10.52601/bpr.2023.220016

Measurement of ATGL activity using adiposomes

doi: 10.52601/bpr.2023.220016
Funds:  The authors would like to thank Dr. Yang Wang for her advices on adiposome preparation and TAG hydrolase assay, Dr. Shimeng Xu for his suggestions on site-directed mutation, and Dr. Liujuan Cui for her help of BAT cytosol extraction. The authors also want to thank Dr. Chang Zhou and Dr. Liujuan Cui for their administrative support. The authors thank Hongjie Zhang for his suggestions of radioisotope experiments. This work was supported the National Natural Science Foundation of China (91857201, 91954108, 32170787 and 32100557) and National Key R&D Program of China (2018YFA0800700 and 2018YFA0800900).
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  • Corresponding author: syzhang@ibp.ac.cn (S. Zhang); pliu@ibp.ac.cn (P. Liu)
  • Received Date: 27 July 2022
  • Accepted Date: 13 January 2023
  • Available Online: 27 March 2023
  • Adipose triacylglycerol lipase (ATGL) is a dynamic lipid droplet-associated protein involved in cellular lipolysis, which is conserved from bacteria to humans. Recent methods that measure the enzymatic activity of ATGL in vitro are established using lipid emulsions. However, the lipid emulsion platforms contain various membranous structures which reduce the accuracy of enzymatic activity determination. Therefore, a new platform and corresponding method are required for accurate measurement of ATGL enzymatic activity that represents cellular lipid and energy homeostasis. Adiposomes are artificial lipid nanostructures mimicking lipid droplets. Employing adiposome as a platform, we have developed an assay to measure the enzymatic activity of ATGL in vitro. Here, a detailed protocol is described to explain how to measure the activity of ATGL using adiposomes. This method successfully proves the concept of lipid droplet-mimetic lipase activity determining platform and provides a tool to identify the active sites of lipases.

  • Xuejing Ma, Zelun Zhi, Shuyan Zhang and Pingsheng Liu declare that they have no conflict of interest.
    All institutional and national guidelines for the care and use of laboratory animals were followed.

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