Diffusion‐based size determination of solute particles a method adapted for postsynaptic proteins /

The postsynaptic density (PSD) is a complex, multilayered protein network largely situated on the internal surface of the postsynaptic membrane. It is the first processing unit for incoming synaptic signals, and changes in its internal structure are associated with synaptic strength and plasticity....

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Bibliographic Details
Main Authors: Szabó András László
Nagy-Kanta Eszter
Varga Soma
Kassainé Jáger Edit
Pongor Csaba István
Laki Mária
Laki András József
Gáspári Zoltán
Format: Article
Published: 2025
Series:FEBS OPEN BIO
Subjects:
doi:10.1002/2211-5463.70111

mtmt:36314112
Online Access:https://publikacio.ppke.hu/2754

MARC

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520 3 |a The postsynaptic density (PSD) is a complex, multilayered protein network largely situated on the internal surface of the postsynaptic membrane. It is the first processing unit for incoming synaptic signals, and changes in its internal structure are associated with synaptic strength and plasticity. These structural changes are largely governed by multivalent interactions between its components. The in vitro characterization of such complexes requires unbiased methods that can be used to estimate the size of the emerging assemblies for systems with multiple possible stoichiometries. Here, we present an experimental method for detecting specific PSD proteins as well as their complexes based on their diffusion in a microfluidic environment. The method requires a fluorescent labeling technique that does not disrupt the function of labeled proteins, a microfluidic device that can maintain laminar flow for protein solutions, a microscope that can record the fluorescent signal emitted by these solutions, and an analytic software package that can process the collected experimental data and convert them into approximate particle sizes. We demonstrate the applicability of our method on protein constructs of various postsynaptic proteins, including the multivalent assembly between GKAP and LC8. 
650 4 |a Biokémia és molekuláris biológia 
700 0 2 |a Nagy-Kanta Eszter  |e aut 
700 0 2 |a Varga Soma  |e aut 
700 0 2 |a Kassainé Jáger Edit  |e aut 
700 0 2 |a Pongor Csaba István  |e aut 
700 0 2 |a Laki Mária  |e aut 
700 0 2 |a Laki András József  |e aut 
700 0 2 |a Gáspári Zoltán  |e aut 
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