Detecting ligand-protein interactions inside cells using reactive peptide tags and split luciferase

2021 ◽  
Author(s):  
Tsuyoshi Takahashi ◽  
Hiroaki Hagiwara
Keyword(s):  
Protein Interactions ◽  
Short Peptide ◽  
Nanoluc Luciferase ◽  
Peptide Tags ◽  
Split Luciferase

We report a method for detecting ligand-protein interactions occurring within cells using short peptide reactive tags appended to ligands and proteins, along with a split NanoLuc luciferase. This method can...

scholarly journals A Global Review on Short Peptides: Frontiers and Perspectives

Molecules ◽  
2021 ◽  
Vol 26 (2) ◽  
pp. 430
Author(s):  
Vasso Apostolopoulos ◽  
Joanna Bojarska ◽  
Tsun-Thai Chai ◽  
Sherif Elnagdy ◽  
Krzysztof Kaczmarek ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Targeted Delivery ◽  
Swot Analysis ◽  
Functional Materials ◽  
Great Promise ◽  
Short Peptides ◽  
Altered Peptide Ligands ◽  
Short Peptide ◽  
Domains Of Life

Peptides are fragments of proteins that carry out biological functions. They act as signaling entities via all domains of life and interfere with protein-protein interactions, which are indispensable in bio-processes. Short peptides include fundamental molecular information for a prelude to the symphony of life. They have aroused considerable interest due to their unique features and great promise in innovative bio-therapies. This work focusing on the current state-of-the-art short peptide-based therapeutical developments is the first global review written by researchers from all continents, as a celebration of 100 years of peptide therapeutics since the commencement of insulin therapy in the 1920s. Peptide “drugs” initially played only the role of hormone analogs to balance disorders. Nowadays, they achieve numerous biomedical tasks, can cross membranes, or reach intracellular targets. The role of peptides in bio-processes can hardly be mimicked by other chemical substances. The article is divided into independent sections, which are related to either the progress in short peptide-based theranostics or the problems posing challenge to bio-medicine. In particular, the SWOT analysis of short peptides, their relevance in therapies of diverse diseases, improvements in (bio)synthesis platforms, advanced nano-supramolecular technologies, aptamers, altered peptide ligands and in silico methodologies to overcome peptide limitations, modern smart bio-functional materials, vaccines, and drug/gene-targeted delivery systems are discussed.

scholarly journals Characterization of Recombinant Flaviviridae Viruses Possessing a Small Reporter Tag

2017 ◽  
Vol 92 (2) ◽  
Author(s):  
Tomokazu Tamura ◽  
Takasuke Fukuhara ◽  
Takuro Uchida ◽  
Chikako Ono ◽  
Hiroyuki Mori ◽  
...  
Keyword(s):  
Life Cycle ◽  
Amino Acid ◽  
Viral Life Cycle ◽  
Luciferase Gene ◽  
Content Type ◽  
Recombinant Viruses ◽  
Nanoluc Luciferase ◽  
The Stability ◽  
Split Luciferase

ABSTRACTThe familyFlaviviridaeconsists of four genera,Flavivirus,Pestivirus,Pegivirus, andHepacivirus, and comprises important pathogens of human and animals. Although the construction of recombinant viruses carrying reporter genes encoding fluorescent and bioluminescent proteins has been reported, the stable insertion of foreign genes into viral genomes retaining infectivity remains difficult. Here, we applied the 11-amino-acid subunit derived from NanoLuc luciferase to the engineering of theFlaviviridaeviruses and then examined the biological characteristics of the viruses. We successfully generated recombinant viruses carrying the split-luciferase gene, including dengue virus, Japanese encephalitis virus, hepatitis C virus (HCV), and bovine viral diarrhea virus. The stability of the viruses was confirmed by five rounds of serial passages in the respective susceptible cell lines. The propagation of the recombinant luciferase viruses in each cell line was comparable to that of the parental viruses. By using a purified counterpart luciferase protein, this split-luciferase assay can be applicable in various cell lines, even when it is difficult to transduce the counterpart gene. The efficacy of antiviral reagents against the recombinant viruses could be monitored by the reduction of luciferase expression, which was correlated with that of viral RNA, and the recombinant HCV was also useful to examine viral dynamicsin vivo. Taken together, our findings indicate that the recombinantFlaviviridaeviruses possessing the split NanoLuc luciferase gene generated here provide powerful tools to understand viral life cycle and pathogenesis and a robust platform to develop novel antivirals againstFlaviviridaeviruses.IMPORTANCEThe construction of reporter viruses possessing a stable transgene capable of expressing specific signals is crucial to investigations of viral life cycle and pathogenesis and the development of antivirals. However, it is difficult to maintain the stability of a large foreign gene, such as those for fluorescence and bioluminescence, after insertion into a viral genome. Here, we successfully generated recombinantFlaviviridaeviruses carrying the 11-amino-acid subunit derived from NanoLuc luciferase and demonstrated that these viruses are applicable toin vitroandin vivoexperiments, suggesting that these recombinantFlaviviridaeviruses are powerful tools for increasing our understanding of viral life cycle and pathogenesis and that these recombinant viruses will provide a robust platform to develop antivirals againstFlaviviridaeviruses.

Analysis and Control of Protein Crystallization Using Short Peptide Tags That Change Solubility without Affecting Structure, Thermal Stability, and Function

2015 ◽  
Vol 15 (6) ◽  
pp. 2703-2711 ◽  
Author(s):  
Mohammad Monirul Islam ◽  
Shigeyoshi Nakamura ◽  
Keiichi Noguchi ◽  
Masafumi Yohda ◽  
Shun-ichi Kidokoro ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Protein Crystallization ◽  
Short Peptide ◽  
Peptide Tags ◽  
And Function ◽  
And Control

Genetically Encoded Short Peptide Tags for Orthogonal Protein Labeling by Sfp and AcpS Phosphopantetheinyl Transferases

2007 ◽  
Vol 2 (5) ◽  
pp. 337-346 ◽  
Author(s):  
Zhe Zhou ◽  
Pablo Cironi ◽  
Alison J. Lin ◽  
Yangqing Xu ◽  
Siniša Hrvatin ◽  
...  
Keyword(s):  
Protein Labeling ◽  
Short Peptide ◽  
Peptide Tags ◽  
Phosphopantetheinyl Transferases

Split Luciferase as an Optical Probe for Detecting Protein−Protein Interactions in Mammalian Cells Based on Protein Splicing

2001 ◽  
Vol 73 (11) ◽  
pp. 2516-2521 ◽  
Author(s):  
Takeaki Ozawa ◽  
Asami Kaihara ◽  
Moritoshi Sato ◽  
Kazunari Tachihara ◽  
Yoshio Umezawa
Keyword(s):  
Protein Interactions ◽  
Mammalian Cells ◽  
Optical Probe ◽  
Protein Splicing ◽  
Protein Protein Interactions ◽  
Split Luciferase

Short peptide tags increase the yield of C-terminally labeled protein

2007 ◽  
Vol 29 (7) ◽  
pp. 1065-1073 ◽  
Author(s):  
Teruaki Kobayashi ◽  
Miwa Shiratori ◽  
Hirofumi Nakano ◽  
Chikashi Eguchi ◽  
Makoto Shirai ◽  
...  
Keyword(s):  
Short Peptide ◽  
Peptide Tags
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