Reconstitution of Membrane-tethered Postsynaptic Density Condensates Using Supported Lipid Bilayer
Eukaryotic cells utilize sub-cellular compartmentalization to restrict reaction components within a defined localization to perform specified biological functions. One way to achieve this is via membrane enclosure; however, many compartments are not bounded with lipid membrane bilayers. In the past few years, it has been increasingly recognized that molecular components in non- or semi-membrane-bound compartments might form biological condensates autonomously (i.e., without requirement of energy input) once threshold concentrations are reached, via a physical chemistry process known as liquid–liquid phase separation. Molecular components within these compartments are stably maintained at high concentrations and separated from the surrounding diluted solution without the need for a physical barrier. Biochemical reconstitution using recombinantly purified proteins has served as an important tool for understanding organizational principles behind these biological condensates. Common techniques include turbidity measurement, fluorescence imaging of 3D droplets, and atomic force microscopy measurements of condensate droplets. Nevertheless, many molecular compartments are semi-membrane-bound with one side attached to the plasma membrane and the other side exposed to the cytoplasm and/or attached to the cytoskeleton; therefore, reconstitution in 3D solution cannot fully recapture their physiological configuration. Here, we utilize a postsynaptic density minimal system to demonstrate that biochemical reconstitution can be applied on supported lipid bilayer (SLB); we have also incorporated actin cytoskeleton into the reconstitution system to mimic the molecular organization in postsynaptic termini. The same system could be adapted to study other membrane-proximal, cytoskeleton-supported condensations.
Ocean Park Conservation Foundation, Hong Kong ; Glaucoma Research Foundation ; Ocean Park Conservation Foundation, Hong Kong ; Ministry of Science and Technology of the People's Republic of China[2019YFA0508402] ; National Natural Science Foundation of China ; Ocean Park Conservation Foundation, Hong Kong[AoE-M09-12] ; Human Frontier Science Program[RGP0020/2019]
Cited Times [WOS]:0
|Document Type||Journal Article|
|Department||School of Life Sciences|
1.State Key Laboratory of Genetic Engineering,School of Life Sciences,Fudan University,Shanghai,200433,China
2.School of Life Sciences,Southern University of Science and Technology,Shenzhen,518055,China
Feng，Zhe,Zhang，Mingjie. Reconstitution of Membrane-tethered Postsynaptic Density Condensates Using Supported Lipid Bilayer[J]. Bio-protocol,2023,13(7).
Feng，Zhe,&Zhang，Mingjie.(2023).Reconstitution of Membrane-tethered Postsynaptic Density Condensates Using Supported Lipid Bilayer.Bio-protocol,13(7).
Feng，Zhe,et al."Reconstitution of Membrane-tethered Postsynaptic Density Condensates Using Supported Lipid Bilayer".Bio-protocol 13.7(2023).
|Files in This Item:||There are no files associated with this item.|
|Recommend this item|
|Export to Endnote|
|Export to Excel|
|Export to Csv|
|Similar articles in Google Scholar|
|Similar articles in Baidu Scholar|
|Similar articles in Bing Scholar|
Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.