In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission Bioprobe
Protein folding is important for protein homeostasis/proteostasis in the human body. We have established the ability to manipulate protein unfolding/refolding for β-lactoglobulin using the induced mechanical energy in the thin film microfluidic vortex fluidic device (VFD) with monitoring as such usi...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-07-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/14/4273 |
| _version_ | 1797526514962530304 |
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| author | Qi Hu Haozhen Hu Xinyi Zhang Kyle Fan Yuning Hong Colin L. Raston Youhong Tang |
| author_facet | Qi Hu Haozhen Hu Xinyi Zhang Kyle Fan Yuning Hong Colin L. Raston Youhong Tang |
| author_sort | Qi Hu |
| collection | DOAJ |
| description | Protein folding is important for protein homeostasis/proteostasis in the human body. We have established the ability to manipulate protein unfolding/refolding for β-lactoglobulin using the induced mechanical energy in the thin film microfluidic vortex fluidic device (VFD) with monitoring as such using an aggregation-induced emission luminogen (AIEgen), TPE-MI. When denaturant (guanidine hydrochloride) is present with β-lactoglobulin, the VFD accelerates the denaturation reaction in a controlled way. Conversely, rapid renaturation of the unfolded protein occurs in the VFD in the absence of the denaturant. The novel TPE-MI reacts with exposed cysteine thiol when the protein unfolds, as established with an increase in fluorescence intensity. TPE-MI provides an easy and accurate way to monitor the protein folding, with comparable results established using conventional circular dichroism. The controlled VFD-mediated protein folding coupled with in situ bioprobe AIEgen monitoring is a viable methodology for studying the denaturing of proteins. |
| first_indexed | 2024-03-10T09:30:53Z |
| format | Article |
| id | doaj.art-caa6b3de6fb847078d5d91e0e33d3837 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T09:30:53Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-caa6b3de6fb847078d5d91e0e33d38372023-11-22T04:31:15ZengMDPI AGMolecules1420-30492021-07-012614427310.3390/molecules26144273In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission BioprobeQi Hu0Haozhen Hu1Xinyi Zhang2Kyle Fan3Yuning Hong4Colin L. Raston5Youhong Tang6Medical Device Research Institute, College of Science and Engineering, Flinders University, Adelaide, SA 5042, AustraliaMedical Device Research Institute, College of Science and Engineering, Flinders University, Adelaide, SA 5042, AustraliaMedical Device Research Institute, College of Science and Engineering, Flinders University, Adelaide, SA 5042, AustraliaFlinders Institute for Nanoscale Science and Technology, College of Science and Engineering, Flinders University, Adelaide, SA 5042, AustraliaDepartment of Chemistry and Physics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC 3086, AustraliaFlinders Institute for Nanoscale Science and Technology, College of Science and Engineering, Flinders University, Adelaide, SA 5042, AustraliaMedical Device Research Institute, College of Science and Engineering, Flinders University, Adelaide, SA 5042, AustraliaProtein folding is important for protein homeostasis/proteostasis in the human body. We have established the ability to manipulate protein unfolding/refolding for β-lactoglobulin using the induced mechanical energy in the thin film microfluidic vortex fluidic device (VFD) with monitoring as such using an aggregation-induced emission luminogen (AIEgen), TPE-MI. When denaturant (guanidine hydrochloride) is present with β-lactoglobulin, the VFD accelerates the denaturation reaction in a controlled way. Conversely, rapid renaturation of the unfolded protein occurs in the VFD in the absence of the denaturant. The novel TPE-MI reacts with exposed cysteine thiol when the protein unfolds, as established with an increase in fluorescence intensity. TPE-MI provides an easy and accurate way to monitor the protein folding, with comparable results established using conventional circular dichroism. The controlled VFD-mediated protein folding coupled with in situ bioprobe AIEgen monitoring is a viable methodology for studying the denaturing of proteins.https://www.mdpi.com/1420-3049/26/14/4273vortex fluidic deviceprotein folding/unfoldingaggregation induced emission |
| spellingShingle | Qi Hu Haozhen Hu Xinyi Zhang Kyle Fan Yuning Hong Colin L. Raston Youhong Tang In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission Bioprobe Molecules vortex fluidic device protein folding/unfolding aggregation induced emission |
| title | In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission Bioprobe |
| title_full | In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission Bioprobe |
| title_fullStr | In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission Bioprobe |
| title_full_unstemmed | In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission Bioprobe |
| title_short | In Situ Monitored Vortex Fluidic-Mediated Protein Refolding/Unfolding Using an Aggregation-Induced Emission Bioprobe |
| title_sort | in situ monitored vortex fluidic mediated protein refolding unfolding using an aggregation induced emission bioprobe |
| topic | vortex fluidic device protein folding/unfolding aggregation induced emission |
| url | https://www.mdpi.com/1420-3049/26/14/4273 |
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