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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Grünberg, Raik Burnier, Julia V. Ferrar, Tony Beltran-Sastre, Violeta Stricher, François Van Der Sloot, Almer M. Garcia-Olivas, Raquel Mallabiabarrena, Arrate Sanjuan, Xavier Zimmermann, Timo Serrano, Luis |
| Description | Country affiliation: Canada Author Affiliation: Grünberg R ( 1] EMBL/CRG Systems Biology Research Unit, Center for Genomic Regulation, Barcelona, Spain. [2] Pompeu Fabra University, Barcelona, Spain. [3] Institut de Recherche en Immunologie et en Cancérologie, Université de Montréal, Montréal, Canada.) |
| Abstract | Fluorescence resonance energy transfer (FRET)-based detection of protein interactions is limited by the very narrow range of FRET-permitting distances. We show two different strategies for the rational design of weak helper interactions that co-recruit donor and acceptor fluorophores for a more robust detection of bimolecular FRET: (i) in silico design of electrostatically driven encounter complexes and (ii) fusion of tunable domain-peptide interaction modules based on WW or SH3 domains. We tested each strategy for optimization of FRET between (m)Citrine and mCherry, which do not natively interact. Both approaches yielded comparable and large increases in FRET efficiencies with little or no background. Helper-interaction modules can be fused to any pair of fluorescent proteins and could, we found, enhance FRET between mTFP1 and mCherry as well as between mTurquoise2 and mCitrine. We applied enhanced helper-interaction FRET (hiFRET) probes to study the binding between full-length H-Ras and Raf1 as well as the drug-induced interaction between Raf1 and B-Raf. |
| File Format | HTM / HTML |
| ISSN | 15487091 |
| Issue Number | 10 |
| Volume Number | 10 |
| e-ISSN | 15487105 |
| Journal | Nature Methods |
| Language | English |
| Publisher | Nature Publishing Group |
| Publisher Date | 2013-10-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Bacterial Proteins Chemistry Fluorescence Resonance Energy Transfer Methods Fluorescent Dyes Luminescent Proteins Protein Interaction Mapping Hela Cells Humans Models, Chemical Models, Molecular Protein Binding Protein Interaction Domains And Motifs Static Electricity Raf Kinases Metabolism Ras Proteins Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology Biotechnology |
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