A split intein and split luciferase-coupled system for detecting protein-protein interactions
Abstract Elucidation of protein-protein interactions (PPIs) represents one of the most important methods in biomedical research. Recently, PPIs have started to be exploited for drug discovery purposes and have thus attracted much attention from both the academic and pharmaceutical sectors. We previo...
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| Format: | Article |
| Language: | English |
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Springer Nature
2024-12-01
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| Series: | Molecular Systems Biology |
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| Online Access: | https://doi.org/10.1038/s44320-024-00081-2 |
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| author | Zhong Yao Jiyoon Kim Betty Geng Jinkun Chen Victoria Wong Anna Lyakisheva Jamie Snider Marina Rudan Dimlić Sanda Raić Igor Stagljar |
| author_facet | Zhong Yao Jiyoon Kim Betty Geng Jinkun Chen Victoria Wong Anna Lyakisheva Jamie Snider Marina Rudan Dimlić Sanda Raić Igor Stagljar |
| author_sort | Zhong Yao |
| collection | DOAJ |
| description | Abstract Elucidation of protein-protein interactions (PPIs) represents one of the most important methods in biomedical research. Recently, PPIs have started to be exploited for drug discovery purposes and have thus attracted much attention from both the academic and pharmaceutical sectors. We previously developed a sensitive method, Split Intein-Mediated Protein Ligation (SIMPL), for detecting binary PPIs via irreversible splicing of the interacting proteins being investigated. Here, we incorporated tripart nanoluciferase (tNLuc) into the system, providing a luminescence signal which, in conjunction with homogenous liquid phase operation, improves the quantifiability and operability of the assay. Using a reference PPI set, we demonstrated an improvement in both sensitivity and specificity over the original SIMPL assay. Moreover, we designed the new SIMPL-tNLuc (‘SIMPL2’) platform with an inherent modularity allowing for flexible measurement of molecular modulators of target PPIs, including inhibitors, molecular glues and PROTACs. Our results demonstrate that SIMPL2 is a sensitive, cost- and labor-effective tool suitable for high-throughput screening (HTS) in both PPI mapping and drug discovery applications. |
| format | Article |
| id | doaj-art-80cffdec304f49ef96ec9948284d6ba6 |
| institution | Kabale University |
| issn | 1744-4292 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-80cffdec304f49ef96ec9948284d6ba62025-02-09T13:00:47ZengSpringer NatureMolecular Systems Biology1744-42922024-12-0121210712510.1038/s44320-024-00081-2A split intein and split luciferase-coupled system for detecting protein-protein interactionsZhong Yao0Jiyoon Kim1Betty Geng2Jinkun Chen3Victoria Wong4Anna Lyakisheva5Jamie Snider6Marina Rudan Dimlić7Sanda Raić8Igor Stagljar9Donnelly Centre, University of TorontoDonnelly Centre, University of TorontoDonnelly Centre, University of TorontoDonnelly Centre, University of TorontoDonnelly Centre, University of TorontoDonnelly Centre, University of TorontoDonnelly Centre, University of TorontoMediterranean Institute for Life Sciences, University of Split School of MedicineMediterranean Institute for Life Sciences, University of Split School of MedicineDonnelly Centre, University of TorontoAbstract Elucidation of protein-protein interactions (PPIs) represents one of the most important methods in biomedical research. Recently, PPIs have started to be exploited for drug discovery purposes and have thus attracted much attention from both the academic and pharmaceutical sectors. We previously developed a sensitive method, Split Intein-Mediated Protein Ligation (SIMPL), for detecting binary PPIs via irreversible splicing of the interacting proteins being investigated. Here, we incorporated tripart nanoluciferase (tNLuc) into the system, providing a luminescence signal which, in conjunction with homogenous liquid phase operation, improves the quantifiability and operability of the assay. Using a reference PPI set, we demonstrated an improvement in both sensitivity and specificity over the original SIMPL assay. Moreover, we designed the new SIMPL-tNLuc (‘SIMPL2’) platform with an inherent modularity allowing for flexible measurement of molecular modulators of target PPIs, including inhibitors, molecular glues and PROTACs. Our results demonstrate that SIMPL2 is a sensitive, cost- and labor-effective tool suitable for high-throughput screening (HTS) in both PPI mapping and drug discovery applications.https://doi.org/10.1038/s44320-024-00081-2Protein-protein InteractionTri-part NanoluciferaseSplit InteinInhibitorDrug Discovery |
| spellingShingle | Zhong Yao Jiyoon Kim Betty Geng Jinkun Chen Victoria Wong Anna Lyakisheva Jamie Snider Marina Rudan Dimlić Sanda Raić Igor Stagljar A split intein and split luciferase-coupled system for detecting protein-protein interactions Molecular Systems Biology Protein-protein Interaction Tri-part Nanoluciferase Split Intein Inhibitor Drug Discovery |
| title | A split intein and split luciferase-coupled system for detecting protein-protein interactions |
| title_full | A split intein and split luciferase-coupled system for detecting protein-protein interactions |
| title_fullStr | A split intein and split luciferase-coupled system for detecting protein-protein interactions |
| title_full_unstemmed | A split intein and split luciferase-coupled system for detecting protein-protein interactions |
| title_short | A split intein and split luciferase-coupled system for detecting protein-protein interactions |
| title_sort | split intein and split luciferase coupled system for detecting protein protein interactions |
| topic | Protein-protein Interaction Tri-part Nanoluciferase Split Intein Inhibitor Drug Discovery |
| url | https://doi.org/10.1038/s44320-024-00081-2 |
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