A split luciferase complementation assay for studying in vivo protein–protein interactions in filamentous ascomycetes

2012 ◽  
Vol 58 (3) ◽  
pp. 179-189 ◽  
Author(s):  
Hee-Kyoung Kim ◽  
Eun Ji Cho ◽  
Seong mi Jo ◽  
Bo Reum Sung ◽  
Seunghoon Lee ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Complementation Assay ◽  
Split Luciferase Complementation ◽  
Split Luciferase

scholarly journals Split luciferase complementation assay to detect regulated protein-protein interactions in rice protoplasts in a large-scale format

Rice ◽  
2014 ◽  
Vol 7 (1) ◽  
Author(s):  
Yukichi Fujikawa ◽  
Takahiro Nakanishi ◽  
Hiroko Kawakami ◽  
Kanako Yamasaki ◽  
Masa H Sato ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Large Scale ◽  
Protein Protein Interactions ◽  
Complementation Assay ◽  
Split Luciferase Complementation ◽  
Scale Format ◽  
Split Luciferase

scholarly journals Application of a split luciferase complementation assay for the detection of viral protein–protein interactions

2011 ◽  
Vol 176 (1-2) ◽  
pp. 108-111 ◽  
Author(s):  
Qiji Deng ◽  
Dan Wang ◽  
Xiaoxiao Xiang ◽  
Xiaofei Gao ◽  
Philip R. Hardwidge ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Viral Protein ◽  
Protein Protein Interactions ◽  
Complementation Assay ◽  
Split Luciferase Complementation ◽  
Split Luciferase

LUCIFERASES FOR THE STUDY OF PROTEIN–PROTEIN INTERACTIONS IN LIVE CELLS AND ANIMALS

Nano LIFE ◽  
2010 ◽  
Vol 01 (01n02) ◽  
pp. 79-87 ◽  
Author(s):  
A. K. M. KAFI ◽  
MITSURU HATTORI ◽  
TAKEAKI OZAWA
Keyword(s):  
Transcriptional Regulation ◽  
Protein Interactions ◽  
Protein Modification ◽  
Specific Protein ◽  
Live Cells ◽  
Protein Protein Interactions ◽  
Recent Developments ◽  
Split Luciferase Complementation ◽  
Spatio Temporal ◽  
Split Luciferase

Many imaging technologies based on luminescent proteins have proven useful for detecting protein–protein interactions, tracking cells in mice, and monitoring transcriptional regulation of specific genes. Especially, novel bioluminescent proteins have advanced the study of induced protein interactions and protein modification in live cells and animals. This review focuses on recent developments of bioluminescent probes for quantitative evaluation of specific protein–protein interactions and their spatio-temporal imaging by means of split luciferase complementation techniques. From the comparison between fluorescent and bioluminescent proteins, advantages and drawbacks of the bioluminescence techniques are described.

scholarly journals Identification of Compounds Targeting Hepatitis B Virus Core Protein Dimerization through a Split Luciferase Complementation Assay

2018 ◽  
Vol 62 (12) ◽  
Author(s):  
Xia-Fei Wei ◽  
Chun-Yang Gan ◽  
Jing Cui ◽  
Ying-Ying Luo ◽  
Xue-Fei Cai ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Protein Interactions ◽  
Core Protein ◽  
Cell Model ◽  
Protein Protein Interactions ◽  
B Virus ◽  
Split Luciferase Complementation ◽  
Split Luciferase

ABSTRACTThe capsid of the hepatitis B virus is an attractive antiviral target for developing therapies against chronic hepatitis B infection. Currently available core protein allosteric modulators (CpAMs) mainly affect one of the two major types of protein-protein interactions involved in the process of capsid assembly, namely, the interaction between the core dimers. Compounds targeting the interaction between two core monomers have not been rigorously screened due to the lack of screening models. We report here a cell-based assay in which the formation of core dimers is indicated by split luciferase complementation (SLC). Making use of this model, 2 compounds, Arbidol (umifenovir) and 20-deoxyingenol, were identified from a library containing 672 compounds as core dimerization regulators. Arbidol and 20-deoxyingenol inhibit the hepatitis B virus (HBV) DNA replicationin vitroby decreasing and increasing the formation of core dimer and capsid, respectively. Our results provided a proof of concept for the cell model to be used to screen new agents targeting the step of core dimer and capsid formation.

Probing the 14-3-3 Isoform-Specificity Profile of Interactions Stabilized by Fusicoccin A

2020 ◽  
Author(s):  
James Frederich ◽  
Ananya Sengupta ◽  
Josue Liriano ◽  
Ewa A. Bienkiewicz ◽  
Brian G. Miller
Keyword(s):  
Protein Interactions ◽  
Neurological Diseases ◽  
Adapter Proteins ◽  
Protein Protein Interactions ◽  
Specific Expression ◽  
Individual Client ◽  
Isoform Specificity ◽  
Fusicoccin A

Fusicoccin A (FC) is a fungal phytotoxin that stabilizes protein–protein interactions (PPIs) between 14-3-3 adapter proteins and their phosphoprotein interaction partners. In recent years, FC has emerged as an important chemical probe of human 14-3-3 PPIs implicated in cancer and neurological diseases. These previous studies have established the structural requirements for FC-induced stabilization of 14-3-3·client phosphoprotein complexes; however, the effect of different 14-3-3 isoforms on FC activity has not been systematically explored. This is a relevant question for the continued development of FC variants because there are seven distinct isoforms of 14-3-3 in humans. Despite their remarkable sequence and structural similarities, a growing body of experimental evidence supports both tissue-specific expression of 14-3-3 isoforms and isoform-specific functions <i>in vivo</i>. Herein, we report the isoform-specificity profile of FC <i>in vitro</i>using recombinant human 14-3-3 isoforms and a focused library of fluorescein-labeled hexaphosphopeptides mimicking the C-terminal 14-3-3 recognition domains of client phosphoproteins targeted by FC in cell culture. Our results reveal modest isoform preferences for individual client phospholigands and demonstrate that FC differentially stabilizes PPIs involving 14-3-3s. Together, these data provide strong motivation for the development of non-natural FC variants with enhanced selectivity for individual 14-3-3 isoforms.

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