AI Article Synopsis
- This study focuses on the interaction between superfolder GFP (sfGFP) and its specific binder nanobody (aGFP), which is important for biomedical research applications involving GFP-tagged proteins.
- The research found that the aGFP nanobody binds strongly to sfGFP, resulting in a significant increase in the stability of aGFP, raising its melting temperature by nearly 30 °C and making the complex stable up to about 85 °C under certain pH conditions.
- These findings indicate that the GFP-aGFP interaction can be effectively used in various applications, including in extreme environments, suggesting its potential for therapeutic uses.
Article Abstract
Objective: The green fluorescent protein (GFP) and its derivatives are widely used in biomedical research. The manipulation of GFP-tagged proteins by GFP-specific binders, e.g. single-domain antibodies (nanobodies), is of increasing significance. It is therefore important to better understand the properties of antiGFP-GFP interaction in order to establish methodological applications. In this work the interaction of superfolder GFP (sfGFP) and its enhancer nanobody (aGFP) was characterized further.
Results: Previous calorimetric experiments demonstrated that the aGFP nanobody binds strongly to sfGFP with a nanomolar affinity. Here we show that this interaction results in a substantial structural stabilization of aGFP reflected in a significant increase of its melting temperature by almost 30 °C. The thermal stability of the sfGFP-aGFP complex is close to 85 °C in the pH range 7.0-8.5. For therapeutic applications thermoresistance is often an essential factor. Our results suggest that methodologies based on GFP-aGFP interaction can be applied under a wide range of physicochemical conditions. The aGFP nanobody seems to be suitable for manipulating sfGFP-labeled targets even in extreme thermophilic organisms.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10283196 | PMC |
| http://dx.doi.org/10.1186/s13104-023-06382-3 | DOI Listing |
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