Multivalent interactions mediated in vitro LLPS
Multivalent interactions are a major driving force behind protein liquid-liquid phase separation. Proteins capable of undergoing LLPS owe their multivalency to the presence of intrinsically disordered regions (IDRs). In this study, we purified a protein with one IDR deleted and one remaining IDR, co...
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sg-ntu-dr.10356-1438902023-02-28T18:07:59Z Multivalent interactions mediated in vitro LLPS Wang, Fengyi Miao Yansong School of Biological Sciences Han Xiao yansongm@ntu.edu.sg Science::Biological sciences::Molecular biology Multivalent interactions are a major driving force behind protein liquid-liquid phase separation. Proteins capable of undergoing LLPS owe their multivalency to the presence of intrinsically disordered regions (IDRs). In this study, we purified a protein with one IDR deleted and one remaining IDR, codenamed TF6Δ-GFP, as well as observed its interactions using microscopy with another protein that contains 2 IDRs, codenamed TF9, and is capable of undergoing LLPS. Our results show that TF6Δ-GFP is unable to undergo LLPS alone and has the tendency to self-aggregate. With the addition of TF9, however, TF6Δ-GFP is recruited into TF9 droplets to form TF9-TF6Δ-GFP condensates. TF6Δ-GFP in these condensates were shown to have liquid-like property by Fluorescence recovery after photobleaching (FRAP) experiments, whereas TF6Δ-GFP aggregates are immobile and do not have liquid-like property. In conclusion, multivalent interactions between TF9 and TF6Δ-GFP allowed TF6Δ-GFP to undergo LLPS as well in vitro despite its self-aggregating nature, which may provide insight as to how proteins that aggregate quickly can still have relevant biological function. Bachelor of Science in Biological Sciences 2020-09-30T01:22:50Z 2020-09-30T01:22:50Z 2020 Final Year Project (FYP) https://hdl.handle.net/10356/143890 en application/pdf Nanyang Technological University |
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Science::Biological sciences::Molecular biology Wang, Fengyi Multivalent interactions mediated in vitro LLPS |
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Multivalent interactions are a major driving force behind protein liquid-liquid phase separation. Proteins capable of undergoing LLPS owe their multivalency to the presence of intrinsically disordered regions (IDRs). In this study, we purified a protein with one IDR deleted and one remaining IDR, codenamed TF6Δ-GFP, as well as observed its interactions using microscopy with another protein that contains 2 IDRs, codenamed TF9, and is capable of undergoing LLPS. Our results show that TF6Δ-GFP is unable to undergo LLPS alone and has the tendency to self-aggregate. With the addition of TF9, however, TF6Δ-GFP is recruited into TF9 droplets to form TF9-TF6Δ-GFP condensates. TF6Δ-GFP in these condensates were shown to have liquid-like property by Fluorescence recovery after photobleaching (FRAP) experiments, whereas TF6Δ-GFP aggregates are immobile and do not have liquid-like property. In conclusion, multivalent interactions between TF9 and TF6Δ-GFP allowed TF6Δ-GFP to undergo LLPS as well in vitro despite its self-aggregating nature, which may provide insight as to how proteins that aggregate quickly can still have relevant biological function. |
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Miao Yansong |
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Miao Yansong Wang, Fengyi |
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Final Year Project |
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Wang, Fengyi |
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Wang, Fengyi |
title |
Multivalent interactions mediated in vitro LLPS |
title_short |
Multivalent interactions mediated in vitro LLPS |
title_full |
Multivalent interactions mediated in vitro LLPS |
title_fullStr |
Multivalent interactions mediated in vitro LLPS |
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Multivalent interactions mediated in vitro LLPS |
title_sort |
multivalent interactions mediated in vitro llps |
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Nanyang Technological University |
publishDate |
2020 |
url |
https://hdl.handle.net/10356/143890 |
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1759856364927057920 |