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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2019
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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. |
format | Online Article Text |
id | pubmed-6921120 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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