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Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body

Protein-Protein Interactions (PPIs) play essential roles in diverse biological processes and their misregulations are associated with a wide range of diseases. Especially, the growing attention to PPIs as a new class of therapeutic target is increasing the need for an efficient method of cell-based...

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Detalles Bibliográficos
Autores principales: Choi, Miri, Baek, Jiyeon, Han, Sang-Bae, Cho, Sungchan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society for Biochemistry and Molecular Biology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235090/
https://www.ncbi.nlm.nih.gov/pubmed/29898808
http://dx.doi.org/10.5483/BMBRep.2018.51.10.051
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author Choi, Miri
Baek, Jiyeon
Han, Sang-Bae
Cho, Sungchan
author_facet Choi, Miri
Baek, Jiyeon
Han, Sang-Bae
Cho, Sungchan
author_sort Choi, Miri
collection PubMed
description Protein-Protein Interactions (PPIs) play essential roles in diverse biological processes and their misregulations are associated with a wide range of diseases. Especially, the growing attention to PPIs as a new class of therapeutic target is increasing the need for an efficient method of cell-based PPI analysis. Thus, we newly developed a robust PPI assay (SeePPI) based on the co-translocation of interacting proteins to the discrete subcellular compartment ‘processing body’ (p-body) inside living cells, enabling a facile analysis of PPI by the enriched fluorescent signal. The feasibility and strength of SeePPI (Signal enhancement exclusively on P-body for Protein-protein Interaction) assay was firmly demonstrated with FKBP12/FRB interaction induced by rapamycin within seconds in real-time analysis of living cells, indicating its recapitulation of physiological PPI dynamics. In addition, we applied p53/MDM2 interaction and its dissociation by Nutlin-3 to SeePPI assay and further confirmed that SeePPI was quantitative and well reflected the endogenous PPI. Our SeePPI assay will provide another useful tool to achieve an efficient analysis of PPIs and their modulators in cells.
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spelling pubmed-62350902018-11-23 Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body Choi, Miri Baek, Jiyeon Han, Sang-Bae Cho, Sungchan BMB Rep Articles Protein-Protein Interactions (PPIs) play essential roles in diverse biological processes and their misregulations are associated with a wide range of diseases. Especially, the growing attention to PPIs as a new class of therapeutic target is increasing the need for an efficient method of cell-based PPI analysis. Thus, we newly developed a robust PPI assay (SeePPI) based on the co-translocation of interacting proteins to the discrete subcellular compartment ‘processing body’ (p-body) inside living cells, enabling a facile analysis of PPI by the enriched fluorescent signal. The feasibility and strength of SeePPI (Signal enhancement exclusively on P-body for Protein-protein Interaction) assay was firmly demonstrated with FKBP12/FRB interaction induced by rapamycin within seconds in real-time analysis of living cells, indicating its recapitulation of physiological PPI dynamics. In addition, we applied p53/MDM2 interaction and its dissociation by Nutlin-3 to SeePPI assay and further confirmed that SeePPI was quantitative and well reflected the endogenous PPI. Our SeePPI assay will provide another useful tool to achieve an efficient analysis of PPIs and their modulators in cells. Korean Society for Biochemistry and Molecular Biology 2018-10 2018-10-31 /pmc/articles/PMC6235090/ /pubmed/29898808 http://dx.doi.org/10.5483/BMBRep.2018.51.10.051 Text en Copyright © 2018 by the The Korean Society for Biochemistry and Molecular Biology This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Choi, Miri
Baek, Jiyeon
Han, Sang-Bae
Cho, Sungchan
Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
title Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
title_full Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
title_fullStr Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
title_full_unstemmed Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
title_short Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
title_sort facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235090/
https://www.ncbi.nlm.nih.gov/pubmed/29898808
http://dx.doi.org/10.5483/BMBRep.2018.51.10.051
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