ArticleInternational journal of molecular sciences2017
A Split-Luciferase Reporter Recognizing GFP and mCherry Tags to Facilitate Studies of Protein-Protein Interactions.
Article in International journal of molecular sciences, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- ATNC: Versatile Nanobody Chimeras for Autophagic Degradation of Intracellular Unligandable and Undruggable Proteins.Journal of the American Chemical Society · 2023Article
- Small molecule-nanobody conjugate induced proximity controls intracellular processes and modulates endogenous unligandable targets.Nature communications · 2023Article
- Expression of functional eGFP-fused antigen-binding fragment of ranibizumab inBioImpacts : BI · 2022Article
- Development of In Vitro and In Vivo Evaluation Systems for Vitamin D Derivatives and Their Application to Drug Discovery.International journal of molecular sciences · 2021Review
- Biophysical Techniques for Target Validation and Drug Discovery in Transcription-Targeted Therapy.International journal of molecular sciences · 2020Review
- Luminescence- and Fluorescence-Based Complementation Assays to Screen for GPCR Oligomerization: Current State of the Art.International journal of molecular sciences · 2019Review
Corrections and comments
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Authors and funding
8 authors.
Funding
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Abstract
The use of fluorescently-tagged proteins in microscopy has become routine, and anti-GFP (Green fluorescent protein) affinity matrices are increasingly used in proteomics protocols. However, some protein-protein interactions assays, such as protein complementation assays (PCA), require recloning of each protein as a fusion with the different parts of the complementation system. Here we describe a generic system where the complementation is separated from the proteins and can be directly used with fluorescently-tagged proteins. By using nanobodies and performing tests in cell-free expression systems, we accelerated the development of multiple reporters, detecting heterodimers and homodimers or oligomers tagged with GFP or mCherry. We demonstrate that the system can detect interactions at a broad range of concentrations, from low nanomolar up to micromolar.
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Registered trials
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