BMB Reports 2018; 51(10): 526-531  https://doi.org/10.5483/BMBRep.2018.51.10.051
Facile analysis of protein-protein interactions in living cells by enriched visualization of the p-body
Miri Choi1,2, Jiyeon Baek1,2, Sang-Bae Han2 and Sungchan Cho1,3,*
1Anticancer Agent Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju 28116, South Korea, 2College of Pharmacy, Chungbuk National University, Cheongju 28644, South Korea, 3Department of Biomolecular Science, KRIBB School of Bioscience, Korea University of Science and Technology, Daejeon 34113, South Korea
Correspondence to: Tel: +82-43-240-6105; Fax: +82-43-240-6159; E-mail: sungchan@kribb.re.kr
Received: March 9, 2018; Revised: April 10, 2018; Accepted: May 25, 2018; Published online: October 31, 2018.
© Korean Society for Biochemistry and Molecular Biology. All rights reserved.

cc 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.
Abstract
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.
Keywords: P-body, PPI modulator, Protein-Protein interaction, Translocation-based PPI assay


This Article


Cited By Articles
  • CrossRef (0)

Funding Information
  • Ministry of Health and Welfare(Ministry of Health, Welfare and Family Affairs)
      10.13039/501100003625
      HI14C2124
  • National Research Foundation of Korea(NRF)
      10.13039/501100003725
      NRF-2016K1A1A8A01938649, NRF2015M3A9C7030128
  • Korea Research Institute of Bioscience and Biotechnology(KRIBB)
      10.13039/501100003715
     

Collections

Services
Social Network Service

e-submission

Archives