A novel system to rapidly detect protein–protein interactions (PPIs) based on fluorescence co-localization
- 13 June 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Biotechnology Letters
- Vol. 42 (11), 2111-2122
- https://doi.org/10.1007/s10529-020-02934-w
Abstract
Objective Rapid and convenient detection of protein–protein interactions (PPIs) is of great significance for understanding function of protein. Results For efficiently detecting PPIs, we used the changes of proteins fluorescence localization to design a novel system, fluorescence translocation co-localization (FTCL), based on nuclear localization signal (NLS) in living cells. Depending on the original state of protein localization (both in the cytoplasm, both in the nucleus, one in the nucleus and another in the cytoplasm), two target proteins can be partitioned into the cytoplasm and nucleus by adding a NLS or mutating an existing NLS. Three independent results display that the changes of protein fluorescence co-localization were observed following co-expression of the two target proteins. At the same time, we verified the accuracy of fluorescence co-localization by co-immunoprecipitation. Conclusions There FTCL system provided a novel detection method for PPIs, regardless of protein localization in the nucleus or cytoplasm. More importantly, this study provides a new strategy for future protein interaction studies through organelle localization (such as mitochondria, Golgi and cytomembrane, etc.).Keywords
Funding Information
- National Natural Science Foundation of China (31872428, 31902214)
- Natural Science Foundation Project of Chongqing, Chongqing Science and Technology Commission (cstc2019jcyj-msxm2371)
- China Agricultural Research System (CARS-18)
This publication has 36 references indexed in Scilit:
- Protein-protein interaction networks: unraveling the wiring of molecular machines within the cellBriefings in Functional Genomics, 2012
- Interactions of the NPXY microdomains of the low density lipoprotein receptor‐related protein 1Proteomics, 2009
- Measurement of FRET Efficiency and Ratio of Donor to Acceptor Concentration in Living CellsBiophysical Journal, 2006
- Mortalin-Based Cytoplasmic Sequestration of p53 in a Nonmammalian Cancer ModelThe American Journal of Pathology, 2006
- Towards a proteome-scale map of the human protein–protein interaction networkNature, 2005
- A Map of the Interactome Network of the Metazoan C. elegansScience, 2004
- A Protein Interaction Map of Drosophila melanogasterScience, 2003
- Comparative assessment of large-scale data sets of protein–protein interactionsNature, 2002
- A novel genetic system to detect protein–protein interactionsNature, 1989
- Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferaseGene, 1988