| Publikationstyp: | Beitrag in wissenschaftlicher Zeitschrift |
| Art der Begutachtung: | Peer review (Publikation) |
| Titel: | Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake |
| Autor/-in: | Chapleau, Richard R. Blomberg, Rebecca Ford, Peter C. Sagermann, Martin |
| DOI: | 10.1110/ps.073358908 |
| Erschienen in: | Protein Science |
| Band(Heft): | 17 |
| Heft: | 4 |
| Seite(n): | 614 |
| Seiten bis: | 622 |
| Erscheinungsdatum: | 2008 |
| Verlag / Hrsg. Institution: | Wiley |
| ISSN: | 0961-8368 1469-896X |
| Sprache: | Englisch |
| Schlagwörter: | Absorption; Binding sites; Biosensing techniques; X-ray crystallography; Escherichia coli; Green Fluorescent Proteins; Mass spectrometry; Mercury; Molecular models; Protein engineering; Fluorescence spectrometry |
| Fachgebiet (DDC): | 660.6: Biotechnologie |
| Zusammenfassung: | Mercury is a ubiquitous pollutant that when absorbed is extremely toxic to a wide variety of biochemical processes. Mercury (II) is a strong, "invisible" poison that is rapidly absorbed by tissues of the intestinal tract, kidneys, and liver upon ingestion. In this study, a novel fluorescence-based biosensor is presented that allows for the direct monitoring of the uptake and distribution of the metal under noninvasive in vivo conditions. With the introduction of a cysteine residue at position 205, located in close proximity to the chromophore, the green fluorescent protein (GFP) from Aequorea victoria was converted into a highly specific biosensor for this metal ion. The mutant protein exhibits a dramatic absorbance and fluorescence change upon mercuration at neutral pH. Absorbance and fluorescence properties with respect to the metal concentration exhibit sigmoidal binding behavior with a detection limit in the low nanomolar range. Time-resolved binding studies indicate rapid subsecond binding of the metal to the protein. The crystal structures obtained of mutant eGFP205C indicate a possible access route of the metal into the core of the protein. To our knowledge, this engineered protein is a first example of a biosensor that allows for noninvasive and real-time imaging of mercury uptake in a living cell. A major advantage is that its expression can be genetically controlled in many organisms to enable unprecedented studies of tissue specific mercury uptake. |
| URI: | https://digitalcollection.zhaw.ch/handle/11475/9666 |
| Volltext Version: | Publizierte Version |
| Lizenz (gemäss Verlagsvertrag): | Lizenz gemäss Verlagsvertrag |
| Departement: | Life Sciences und Facility Management |
| Organisationseinheit: | Institut für Chemie und Biotechnologie (ICBT) |
| Enthalten in den Sammlungen: | Publikationen Life Sciences und Facility Management |
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Chapleau, R. R., Blomberg, R., Ford, P. C., & Sagermann, M. (2008). Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake. Protein Science, 17(4), 614–622. https://doi.org/10.1110/ps.073358908
Chapleau, R.R. et al. (2008) ‘Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake’, Protein Science, 17(4), pp. 614–622. Available at: https://doi.org/10.1110/ps.073358908.
R. R. Chapleau, R. Blomberg, P. C. Ford, and M. Sagermann, “Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake,” Protein Science, vol. 17, no. 4, pp. 614–622, 2008, doi: 10.1110/ps.073358908.
CHAPLEAU, Richard R., Rebecca BLOMBERG, Peter C. FORD und Martin SAGERMANN, 2008. Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake. Protein Science. 2008. Bd. 17, Nr. 4, S. 614–622. DOI 10.1110/ps.073358908
Chapleau, Richard R., Rebecca Blomberg, Peter C. Ford, and Martin Sagermann. 2008. “Design of a Highly Specific and Noninvasive Biosensor Suitable for Real-Time in Vivo Imaging of Mercury (II) Uptake.” Protein Science 17 (4): 614–22. https://doi.org/10.1110/ps.073358908.
Chapleau, Richard R., et al. “Design of a Highly Specific and Noninvasive Biosensor Suitable for Real-Time in Vivo Imaging of Mercury (II) Uptake.” Protein Science, vol. 17, no. 4, 2008, pp. 614–22, https://doi.org/10.1110/ps.073358908.
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