|Publication type:||Article in scientific journal|
|Type of review:||Peer review (publication)|
|Title:||Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake|
|Authors:||Chapleau, Richard R.|
Ford, Peter C.
|Published in:||Protein Science|
|Publisher / Ed. Institution:||Wiley|
|Subjects:||Absorption; Binding sites; Biosensing techniques; X-ray crystallography; Escherichia coli; Green Fluorescent Proteins; Mass spectrometry; Mercury; Molecular models; Protein engineering; Fluorescence spectrometry|
|Subject (DDC):||660.6: Biotechnology|
|Abstract:||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.|
|Fulltext version:||Published version|
|License (according to publishing contract):||Licence according to publishing contract|
|Departement:||Life Sciences and Facility Management|
|Organisational Unit:||Institute of Chemistry and Biotechnology (ICBT)|
|Appears in collections:||Publikationen Life Sciences und Facility Management|
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