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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| Format: | Final Year Project (FYP) |
| Language: | English |
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Nanyang Technological University
2020
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| Online Access: | https://hdl.handle.net/10356/143890 |
| _version_ | 1811691741382705152 |
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| author | Wang, Fengyi |
| author2 | Miao Yansong |
| author_facet | Miao Yansong Wang, Fengyi |
| author_sort | Wang, Fengyi |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T06:24:42Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/143890 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:24:42Z |
| publishDate | 2020 |
| publisher | Nanyang Technological University |
| record_format | dspace |
| spelling | ntu-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 |
| spellingShingle | Science::Biological sciences::Molecular biology Wang, Fengyi Multivalent interactions mediated in vitro LLPS |
| title | Multivalent interactions mediated in vitro LLPS |
| title_full | Multivalent interactions mediated in vitro LLPS |
| title_fullStr | Multivalent interactions mediated in vitro LLPS |
| title_full_unstemmed | Multivalent interactions mediated in vitro LLPS |
| title_short | Multivalent interactions mediated in vitro LLPS |
| title_sort | multivalent interactions mediated in vitro llps |
| topic | Science::Biological sciences::Molecular biology |
| url | https://hdl.handle.net/10356/143890 |
| work_keys_str_mv | AT wangfengyi multivalentinteractionsmediatedinvitrollps |