Measurement of ATGL activity using adiposomes

Ma Xuejing; Zhi Zelun; Zhang Shuyan; Liu Pingsheng

doi:10.52601/bpr.2023.220016

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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

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

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Measurement of ATGL activity using adiposomes

doi: 10.52601/bpr.2023.220016

Ma Xuejing^{1, 2, 3},
Zhi Zelun¹,
Zhang Shuyan^{1, 4, 5
,
,},
Liu Pingsheng^{1, 2
,
,}

1.
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
2.
University of Chinese Academy of Sciences, Beijing 100049, China
3.
Department of Life Sciences, Cangzhou Normal University, Cangzhou 061001, Hebei, China
4.
Institute of Infectious Diseases, Beijing Key Laboratory of Emerging Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China
5.
Beijing Institute of Infectious Diseases, Beijing 100015, China

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).

More Information

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

Abstract

Abstract

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.
- ATGL,
- Enzymatic activity,
- Adiposome,
- Lipid droplet

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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