Title: Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake
Authors : Chapleau, Richard R.
Blomberg, Rebecca
Ford, Peter C.
Sagermann, Martin
Published in : Protein science
Volume(Issue) : 17
Issue : 4
Pages : 614
Pages to: 622
Publisher / Ed. Institution : Wiley
Issue Date: 2008
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (Publication)
Language : English
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.
Departement: Life Sciences and Facility Management
Organisational Unit: Institute of Chemistry and Biotechnology (ICBT)
Publication type: Article in scientific Journal
DOI : 10.1110/ps.073358908
ISSN: 0961-8368
1469-896X
URI: https://digitalcollection.zhaw.ch/handle/11475/9666
Appears in Collections:Publikationen Life Sciences und Facility Management

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