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

Szabó András László; Nagy-Kanta Eszter; Varga Soma; Kassainé Jáger Edit; Pongor Csaba István; Laki Mária; Laki András József; Gáspári Zoltán: Diffusion‐based size determination of solute particles: a method adapted for postsynaptic proteins.
FEBS OPEN BIO. pp. 1-16. ISSN 2211-5463 (2025) (Benyújtva)

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Folyóiratcikk

Szerző azonosítók:

Név	ORCID	MTMT szerző azonosító
Szabó András László	0000-0001-7594-3087	10074260
Nagy-Kanta Eszter	0009-0003-1322-2008	10065218
Varga Soma		10079631
Kassainé Jáger Edit	0000-0003-2601-8391	10002870
Pongor Csaba István	0000-0002-6820-1285	10036100
Laki Mária	0009-0001-4760-6347	10100826
Laki András József		10029874
Gáspári Zoltán	0000-0002-8692-740X	10001271

Absztrakt (kivonat):

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.

Folyóirat címe:

FEBS OPEN BIO

Megjelenés éve:

2025

Oldalak:

pp. 1-16

ISSN:

2211-5463

Intézmény:

Pázmány Péter Katolikus Egyetem

Kar:

Információs Technológiai és Bionikai Kar (2013.07.-)