[Protein complementation as tool for studying protein-protein interactions in living cells]
- PMID: 23156669
[Protein complementation as tool for studying protein-protein interactions in living cells]
Abstract
Association and degradation of protein complexes play essential role in a majority of normal and pathologic processes, which take place in living cell. Studying the underlying mechanisms of those interactions would give deeper understanding of specific causes of disease progression and would allow developing new therapeutic strategies. The majority of technical approaches currently used for detecting protein association include in vitro protein extraction and purification, whereas more relevant results require methods that can be used in vivo. One of a few approaches for in vivo protein association detection is based on reporter protein fragment complementation. Reporter systems based on protein complementation rely on reconstitution of reporter protein fluorescent or enzymatic activity which occurs upon reassociation of protein fragments and could be measured by colorimetry, luminometry or fluorimetry. Protein complementation is widely used to develop reporter systems for analysis of protein interactions, for functional dissection of signal transduction pathways and for performing high-throughput screenings to discover new protein interaction partners. Currently developed approaches that utilize protein fragment complementation have possibilities that extend far beyond simple detection of interaction in a pair of proteins.
Similar articles
-
Protein-fragment complementation assays for large-scale analysis, functional dissection and dynamic studies of protein-protein interactions in living cells.Methods Mol Biol. 2011;756:395-425. doi: 10.1007/978-1-61779-160-4_25. Methods Mol Biol. 2011. PMID: 21870242
-
Using the beta-lactamase protein-fragment complementation assay to probe dynamic protein-protein interactions.Nat Protoc. 2007;2(9):2302-6. doi: 10.1038/nprot.2007.356. Nat Protoc. 2007. PMID: 17853887
-
Detecting Protein-Protein Interaction Based on Protein Fragment Complementation Assay.Curr Protein Pept Sci. 2020;21(6):598-610. doi: 10.2174/1389203721666200213102829. Curr Protein Pept Sci. 2020. PMID: 32053071 Review.
-
Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system.Anal Biochem. 2014 Mar 1;448:92-4. doi: 10.1016/j.ab.2013.12.009. Epub 2013 Dec 11. Anal Biochem. 2014. PMID: 24333278
-
Current state of imaging protein-protein interactions in vivo with genetically encoded reporters.Annu Rev Biomed Eng. 2007;9:321-49. doi: 10.1146/annurev.bioeng.9.060906.152044. Annu Rev Biomed Eng. 2007. PMID: 17461729 Review.
Cited by
-
Surveying the landscape of optogenetic methods for detection of protein-protein interactions.Wiley Interdiscip Rev Syst Biol Med. 2018 May;10(3):e1415. doi: 10.1002/wsbm.1415. Epub 2018 Jan 15. Wiley Interdiscip Rev Syst Biol Med. 2018. PMID: 29334187 Free PMC article. Review.
-
Super-resolution imaging and tracking of protein-protein interactions in sub-diffraction cellular space.Nat Commun. 2014 Jul 17;5:4443. doi: 10.1038/ncomms5443. Nat Commun. 2014. PMID: 25030837 Free PMC article.
-
Directed Evolution of Split APEX2 Peroxidase.ACS Chem Biol. 2019 Apr 19;14(4):619-635. doi: 10.1021/acschembio.8b00919. Epub 2019 Mar 8. ACS Chem Biol. 2019. PMID: 30848125 Free PMC article.