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Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform

New strategies are urgently needed to characterize the functions of the lipid droplet (LD). Here, adiposome, an artificial LD mimetic platform, was validated by comparative in vitro bioassays. Scatchard analysis found that the binding of perilipin 2 (PLIN2) to the adiposome surface was saturable. Ph...

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Autores principales: Ma, Xuejing, Zhi, Zelun, Zhang, Shuyan, Zhou, Chang, Mechler, Adam, Liu, Pingsheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8326204/
https://www.ncbi.nlm.nih.gov/pubmed/34368652
http://dx.doi.org/10.1016/j.isci.2021.102834
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author Ma, Xuejing
Zhi, Zelun
Zhang, Shuyan
Zhou, Chang
Mechler, Adam
Liu, Pingsheng
author_facet Ma, Xuejing
Zhi, Zelun
Zhang, Shuyan
Zhou, Chang
Mechler, Adam
Liu, Pingsheng
author_sort Ma, Xuejing
collection PubMed
description New strategies are urgently needed to characterize the functions of the lipid droplet (LD). Here, adiposome, an artificial LD mimetic platform, was validated by comparative in vitro bioassays. 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 73(rd) glutamic acid of PLIN2 is significant for the effect of PtdIns on the PLIN2 binding. Furthermore, adiposome was also found to be an ideal platform for in situ enzymatic 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 proteins in vitro.
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spelling pubmed-83262042021-08-06 Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform Ma, Xuejing Zhi, Zelun Zhang, Shuyan Zhou, Chang Mechler, Adam Liu, Pingsheng iScience Article New strategies are urgently needed to characterize the functions of the lipid droplet (LD). Here, adiposome, an artificial LD mimetic platform, was validated by comparative in vitro bioassays. 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 73(rd) glutamic acid of PLIN2 is significant for the effect of PtdIns on the PLIN2 binding. Furthermore, adiposome was also found to be an ideal platform for in situ enzymatic 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 proteins in vitro. Elsevier 2021-07-10 /pmc/articles/PMC8326204/ /pubmed/34368652 http://dx.doi.org/10.1016/j.isci.2021.102834 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Ma, Xuejing
Zhi, Zelun
Zhang, Shuyan
Zhou, Chang
Mechler, Adam
Liu, Pingsheng
Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform
title Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform
title_full Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform
title_fullStr Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform
title_full_unstemmed Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform
title_short Validating an artificial organelle: Studies of lipid droplet-specific proteins on adiposome platform
title_sort validating an artificial organelle: studies of lipid droplet-specific proteins on adiposome platform
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8326204/
https://www.ncbi.nlm.nih.gov/pubmed/34368652
http://dx.doi.org/10.1016/j.isci.2021.102834
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