Protocol for using artificial lipid droplets to study the binding affinity of lipid droplet-associated proteins.
Cell Biology
Cell Membrane
Protein expression and purification
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
18 03 2022
18 03 2022
Historique:
entrez:
10
3
2022
pubmed:
11
3
2022
medline:
23
4
2022
Statut:
epublish
Résumé
Here, we present a protocol to construct artificial lipid droplets to study the binding affinity of lipid droplet-associated proteins. We provide procedures to construct adiposomes and prepare recombinant lipid droplet-associated proteins. Then we describe approaches to measure the number density of perilipin 2 on natural lipid droplets, construct artificial lipid droplets, and determine the binding affinity of perilipin 2 on artificial lipid droplets. This protocol can be adapted to determine the binding properties of various lipid droplet-associated proteins. For complete details on the use and execution of this protocol, please refer to Ma et al. (2021).
Identifiants
pubmed: 35265861
doi: 10.1016/j.xpro.2022.101214
pii: S2666-1667(22)00094-6
pmc: PMC8899027
doi:
Substances chimiques
Lipid Droplet Associated Proteins
0
Perilipin-1
0
Perilipin-2
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
101214Informations de copyright
© 2022 The Author(s).
Déclaration de conflit d'intérêts
P.L., X.M., Z.Z., and C.Z. have been authorized to hold a patent filed for the use of artificial LD and application thereof in evaluating interaction between LD-associated proteins and LDs. The interest has been fully disclosed to the Institute of Biophysics, Chinese Academy of Sciences. All other authors declare no competing interests.
Références
Biochim Biophys Acta Mol Cell Biol Lipids. 2017 Oct;1862(10 Pt B):1166-1177
pubmed: 28627435
Nat Commun. 2017 Jul 06;8:15979
pubmed: 28681845
J Lipid Res. 2004 Nov;45(11):1983-91
pubmed: 15342676
J Biol Chem. 2004 Jan 30;279(5):3787-92
pubmed: 14597625
Anal Biochem. 1967 Sep;20(3):525-32
pubmed: 6048188
Plant Physiol. 2016 Apr;170(4):2052-71
pubmed: 26896396
iScience. 2021 Jul 10;24(8):102834
pubmed: 34368652
Biochim Biophys Acta Biomembr. 2021 Aug 1;1863(8):183624
pubmed: 33933429
Cold Spring Harb Perspect Biol. 2011 Mar 01;3(3):
pubmed: 21421923
Nat Protoc. 2013 Jan;8(1):43-51
pubmed: 23222457
Semin Cell Dev Biol. 2020 Dec;108:4-13
pubmed: 32201131
Nat Methods. 2012 Jul;9(7):671-5
pubmed: 22930834
ACS Nano. 2016 Mar 22;10(3):3312-22
pubmed: 26910792