摘要：SummaryNew strategies are urgently needed to characterize the functions of the lipid droplet (LD). Here, adiposome, an artificial LD mimetic platform, was validated by comparativein vitrobioassays. Scatchard analysis found that the binding of perilipin 2 (PLIN2) to the adiposome surface was saturable. Phosphatidylinositol (PtdIns) was found to inhibit PLIN2 binding while it did not impede perilipin 3 (PLIN3). Structural analysis combined with mutagenesis revealed that the 73rdglutamic acid of PLIN2 is significant for the effect of PtdIns on the PLIN2 binding. Furthermore, adiposome was also found to be an ideal platform forin situenzymatic activity measurement of adipose triglyceride lipase (ATGL). The significant serine mutants of ATGL were found to cause the loss of lipase activity. Our study demonstrates the adiposome as a powerful, manipulatable model system that mimics the function of LD for binding and enzymatic activity studies of LD proteinsin vitro.Graphical abstractDisplay OmittedHighlights•An artificial organelle was validated for the targeting and regulation of LD proteins•Binding affinity analysis of PLIN2 targeting on artificial LD was performed•PtdIns and the 73rdglutamic acid in PLIN2 affect the targeting of PLIN2 to LD•In situenzymatic activity of ATGL was demonstrated and measured via artificial LDCell biology; Functional aspects of cell biology; Biophysics
