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A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation

Liquid-liquid phase separation (LLPS) phenomena have been observed in vitro as well as in vivo and came in focus of interdisciplinary research activities particularly aiming at understanding the physico-chemical pathways of LLPS and its functionality in recent years. Dynamic light scattering (DLS) h...

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Autores principales: Falke, Sven, Brognaro, Hévila, Martirosyan, Arayik, Dierks, Karsten, Betzel, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6921120/
https://www.ncbi.nlm.nih.gov/pubmed/31886430
http://dx.doi.org/10.1016/j.heliyon.2019.e03016
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author Falke, Sven
Brognaro, Hévila
Martirosyan, Arayik
Dierks, Karsten
Betzel, Christian
author_facet Falke, Sven
Brognaro, Hévila
Martirosyan, Arayik
Dierks, Karsten
Betzel, Christian
author_sort Falke, Sven
collection PubMed
description Liquid-liquid phase separation (LLPS) phenomena have been observed in vitro as well as in vivo and came in focus of interdisciplinary research activities particularly aiming at understanding the physico-chemical pathways of LLPS and its functionality in recent years. Dynamic light scattering (DLS) has been proven to be a most efficient method to analyze macromolecular clustering in solutions and suspensions with diverse applications in life sciences, material science and biotechnology. For spatially and time-resolved investigations of LLPS, i.e. formation of liquid dense protein clusters (LDCs) and aggregation, a novel eight-channel in situ DLS instrument was designed, constructed and applied. The real time formation of LDCs of glucose isomerase (GI) and bovine pancreatic trypsin inhibitor (BPTI) under different physico-chemical conditions was investigated in situ. Complex shifts in the particle size distributions indicated growth of LDCs up to the μm size regime. Additionally, near-UV circular dichroism spectroscopy was performed to monitor the folding state of the proteins in the process of LDC formation.
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spelling pubmed-69211202019-12-27 A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation Falke, Sven Brognaro, Hévila Martirosyan, Arayik Dierks, Karsten Betzel, Christian Heliyon Article Liquid-liquid phase separation (LLPS) phenomena have been observed in vitro as well as in vivo and came in focus of interdisciplinary research activities particularly aiming at understanding the physico-chemical pathways of LLPS and its functionality in recent years. Dynamic light scattering (DLS) has been proven to be a most efficient method to analyze macromolecular clustering in solutions and suspensions with diverse applications in life sciences, material science and biotechnology. For spatially and time-resolved investigations of LLPS, i.e. formation of liquid dense protein clusters (LDCs) and aggregation, a novel eight-channel in situ DLS instrument was designed, constructed and applied. The real time formation of LDCs of glucose isomerase (GI) and bovine pancreatic trypsin inhibitor (BPTI) under different physico-chemical conditions was investigated in situ. Complex shifts in the particle size distributions indicated growth of LDCs up to the μm size regime. Additionally, near-UV circular dichroism spectroscopy was performed to monitor the folding state of the proteins in the process of LDC formation. Elsevier 2019-12-13 /pmc/articles/PMC6921120/ /pubmed/31886430 http://dx.doi.org/10.1016/j.heliyon.2019.e03016 Text en © 2019 The Author(s) http://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
Falke, Sven
Brognaro, Hévila
Martirosyan, Arayik
Dierks, Karsten
Betzel, Christian
A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation
title A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation
title_full A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation
title_fullStr A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation
title_full_unstemmed A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation
title_short A multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation
title_sort multi-channel in situ light scattering instrument utilized for monitoring protein aggregation and liquid dense cluster formation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6921120/
https://www.ncbi.nlm.nih.gov/pubmed/31886430
http://dx.doi.org/10.1016/j.heliyon.2019.e03016
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