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A novel system to rapidly detect protein–protein interactions (PPIs) based on fluorescence co-localization

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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.).

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (31872428 and 31902214), Natural Science Foundation of Chongqing (cstc2019jcyj-msxm2371) and the China Agriculture Research System (CARS-18).

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Author notes

  1. Nan Hu and Zhan-Qi Dong have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China

    Nan Hu, Zhan-Qi Dong, Ting-Ting Chen, Ning Zheng, Qin Wu, Peng Chen, Cheng Lu & Min-Hui Pan

  2. Key Laboratory of Sericultural Biology and Genetic Breeding, Ministry of Agriculture, Southwest University, Chongqing, 400716, China

    Zhan-Qi Dong, Peng Chen, Cheng Lu & Min-Hui Pan

Authors
  1. Nan Hu
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  2. Zhan-Qi Dong
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  3. Ting-Ting Chen
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  4. Ning Zheng
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  5. Qin Wu
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  6. Peng Chen
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  7. Cheng Lu
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  8. Min-Hui Pan
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Corresponding authors

Correspondence to Cheng Lu or Min-Hui Pan.

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This article does not contain any experiments with human participants or animals (except invertebrate’s cell lines, which are exempt from ethical concerns) performed by any of the authors.

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10529_2020_2934_MOESM1_ESM.doc

Figure S1: The functional domain of LEF-11. Large rectangle shows the amino acid sequence of LEF-11 where gray areas represent functional motifs and black rectangles represent different LEF-11 forms. The nuclear localization signal is located at the C-terminal, and mutation of the 100th amino acid residue K influences LEF-11 nuclear localization. (DOC 75 kb)

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Hu, N., Dong, ZQ., Chen, TT. et al. A novel system to rapidly detect protein–protein interactions (PPIs) based on fluorescence co-localization. Biotechnol Lett 42, 2111–2122 (2020). https://doi.org/10.1007/s10529-020-02934-w

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