Measurement of ATGL activity using adiposomes

Xuejing Ma; Zelun Zhi; Shuyan Zhang; Pingsheng Liu

doi:10.52601/bpr.2023.220016

Measurement of ATGL activity using adiposomes

Biophys Rep. 2023 Feb 28;9(1):3-14. doi: 10.52601/bpr.2023.220016.

Authors

Xuejing Ma^{1

2

3}, Zelun Zhi¹, Shuyan Zhang^{1

4

5}, Pingsheng Liu^{1

2}

Affiliations

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.

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.

Keywords: ATGL; Adiposome; Enzymatic activity; Lipid droplet.