A new bivalent fluorescent fusion protein for differential Cu(II) and Zn(II) ion detection in aqueous solution

Sorenson, A.E., and Schaeffer, P.M. (2020) A new bivalent fluorescent fusion protein for differential Cu(II) and Zn(II) ion detection in aqueous solution. Analytica Chimica Acta, 1101. pp. 120-128.

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Abstract

Simple and easy to engineer metal-sensing molecules that are capable of differentiating metal ions and producing metal-specific signals are highly desirable. Metal ions affect the thermal stability of proteins by increasing or decreasing their resistance to unfolding. This work illustrates a new strategy for designing bivalent fluorescent fusion proteins capable of differentiating metal ions in solution through their distinct effects on a protein's thermal stability. A new dual purpose metal sensor was developed consisting of biotin protein ligase (BirA) from B. pseudomallei (Bp) fused to green fluorescent protein (GFP). When coupled with differential scanning fluorimetry of GFP-tagged proteins (DSF-GTP) for signal-transduction detection, Bp BirA-GFP yields distinct protein unfolding signatures with Zn(II) and Cu(II) ions in aqueous solutions. The limit of detection of the system is ∼1 μM for both metal species. The system can be used in a variety of high-throughput assay formats including for the screening of metal-binding proteins and chelators. Bp BirA-GFP has also the additional benefit of being useful in Cu(II) ion field-testing applications through simple visual observation of a temperature-dependent loss of fluorescence. Bp BirA-GFP is the first example of a 2protein-based dual purpose Cu(II) and Zn(II) ion sensor compatible with two different yet complementary signal-transduction detection systems.

Item ID: 61890
Item Type: Article (Research - C1)
ISSN: 1873-4324
Keywords: Biosensor, Biotin protein ligase, Burkholderia, Chemodosimeter, Copper, Dengue RNA-Dependent-RNA-Polymerase, Differential scanning fluorimetry, DNA ligase, DnaG primase, DSF-GTP, Green fluorescent protein, HIV integrase, Mycobacterium, Zinc
Copyright Information: © 2019 Elsevier B.V.
Date Deposited: 27 Jan 2021 04:18
FoR Codes: 34 CHEMICAL SCIENCES > 3401 Analytical chemistry > 340108 Sensor technology (incl. chemical aspects) @ 100%
SEO Codes: 96 ENVIRONMENT > 9611 Physical and Chemical Conditions of Water > 961101 Physical and Chemical Conditions of Water for Urban and Industrial Use @ 25%
96 ENVIRONMENT > 9611 Physical and Chemical Conditions of Water > 961102 Physical and Chemical Conditions of Water in Coastal and Estuarine Environments @ 25%
96 ENVIRONMENT > 9611 Physical and Chemical Conditions of Water > 961103 Physical and Chemical Conditions of Water in Fresh, Ground and Surface Water Environments (excl. Urban and @ 50%
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