A General Strategy to Convert the MerR Family Proteins into Highly Sensitive and Selective Fluorescent Biosensors for Metal Ions

Chen, Peng; He, Chuan

doi:10.1021/ja0383975

Cited by 274 publications

(151 citation statements)

References 23 publications

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“…metalloregulatory proteins [17] or metallothioneins [18]) or pre-designed oligopeptides fused to carrier proteins [14,15,[19][20][21][22][23] may be promising candidates in metal ion removal. Appropriate peptides [6,[24][25][26] or the highly efficient and selective bacterial metal responsive metalloregulatory proteins [27] are also utilized in the construction of biosensors [7,8,[28][29][30] for the detection of toxic metal ions. The applied sequences generally contain cysteines with a soft sulfur donor atom showing high affinity 4 towards the heavy metal ions.…”

Section: Introductionmentioning

confidence: 99%

Competition of zinc(II) with cadmium(II) or mercury(II) in binding to a 12-mer peptide

Jancsó

Gyurcsik

Mesterházy

et al. 2013

Journal of Inorganic Biochemistry

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Section: Introductionmentioning

confidence: 99%

Competition of zinc(II) with cadmium(II) or mercury(II) in binding to a 12-mer peptide

Jancsó

Gyurcsik

Mesterházy

et al. 2013

Journal of Inorganic Biochemistry

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“…An alternative approach towards mercury detection is to develop portable sensors. [1] Such sensors have been designed for mercury detection in water using small molecule chelators, [6][7][8] proteins, [9,10] conjugated polymers, [11] nanoparticles, [12] and genetically modified cells. [13] Some of the sensors have been used for the detection of mercury in environmental and biological samples.…”

Section: Introductionmentioning

confidence: 99%

Immobilization of DNA on Magnetic Microparticles for Mercury Enrichment and Detection with Flow Cytometry

Huang

Liu

2011

Chemistry A European J

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Mercury detection in water has attracted a lot of research interest due to its highly toxic nature and adverse environmental impact. In particular, the recent discovery of specific binding of Hg (II) to thyminerich DNA resulting in T-Hg (II) -T base pairs has led to the development of a number of sensors with different signaling mechanisms. However, majority of such sensors were nonimmobilized. Immobilization, on the other hand, allows active mercury adsorption, signal amplification, and sensor regeneration. In this work, we immobilized a thymine-rich DNA on a magnetic microparticle surface via biotin-streptavidin interactions. In the presence of Hg (II) , the DNA changes from a random coil structure into a hairpin, upon which SYBR Green I binds to emit green fluorescence. Detection was carried out using flow cytometry where fluorescence intensity increased ~9-fold in the presence of mercury and the binding of mercury reached equilibrium in less than 2 min. The sensor showed a unique samplevolume dependent fluorescence signal change where a higher fluorescence was obtained with a larger sample volume, suggesting that the particles can actively adsorb Hg (II) . Detection limits of 5 nM (1 ppb) and 14 nM (2.8 ppb) were achieved in pure buffer and in mercury spiked Lake Ontario water samples, respectively.

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“…1A). Its 2¢-deoxy ribose variant has been successfully used, for example, in the characterization of the transcription bubble in elongation complexes of T7 RNA polymerase (Liu andMartin 2001, 2002), in a biosensor for metal ions (Chen and He 2004), and to characterize local structure in a double-stranded DNA (Johnson et al 2005) or DNA/ RNA hybrid (Dash et al 2004). Its unperturbed WatsonCrick face allows PC to base-pair specifically with G, similar to an unmodified cytidine (Fig.…”

Section: Introductionmentioning

confidence: 99%

“…PC is highly fluorescent, with excitation and emission maxima of 350 and 460 nm, respectively, relatively far from those of RNA and protein, making it a suitable probe for both RNAs and RNA-protein complexes. Furthermore, the fluorescence of PC is significantly quenched in duplex DNA and RNA, most likely through base-stacking interactions with surrounding nucleotides, rendering it sensitive to its local environment (Liu andMartin 2001, 2002;Berry et al 2004;Chen and He 2004;Dash et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Pyrrolo-C as a fluorescent probe for monitoring RNA secondary structure formation

Tinsley¹,

Walter²

2006

RNA

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Pyrrolo-C (PC), or 3-[b-D-2-ribofuranosyl]-6-methylpyrrolo [2,3-d]pyrimidin-2(3H)-one, is a fluorescent analog of the nucleoside cytidine that retains its Watson-Crick base-pairing capacity with G. Due to its red-shifted absorbance, it can be selectively excited in the presence of natural nucleosides, making it a potential site-specific probe for RNA structure and dynamics. Similar to 2-aminopurine nucleoside, which base-pairs with uridine (or thymidine), PC's fluorescence becomes reversibly quenched upon base-pairing, most likely due to stacking interactions with neighboring bases. To test its utility as an RNA probe, we examined PC's fluorescent properties over a wide range of ionic strengths, pH, organic cosolvents, and temperatures. Incorporation of PC into a single-stranded RNA results in an $60% reduction of fluorescence intensity, while duplex formation reduces the fluorescence by $75% relative to the free ribonucleoside. We find that the fluorescence intensity of PC is only moderately affected by ionic strength, pH, and temperature, while it is slightly enhanced by organic cosolvents, making it a versatile probe for a broad range of buffer conditions. We demonstrate two applications for PC: fluorescent measurements of the kinetics of formation and dissociation of an RNA/DNA complex, and fluorescent monitoring of the thermal denaturation of the central segment of an RNA duplex. Taken together, our data showcase the potential of pyrrolo-C as an effective fluorescent probe to study RNA structure, dynamics, and function, complementary to the popular 2-aminopurine ribonucleoside.

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A General Strategy to Convert the MerR Family Proteins into Highly Sensitive and Selective Fluorescent Biosensors for Metal Ions

Abstract: The MerR family metal-regulatory proteins are converted into fluorescent biosensors that can detect different metal ions with high sensitivity and selectivity.

Cited by 274 publications

References 23 publications

Competition of zinc(II) with cadmium(II) or mercury(II) in binding to a 12-mer peptide

Competition of zinc(II) with cadmium(II) or mercury(II) in binding to a 12-mer peptide

Immobilization of DNA on Magnetic Microparticles for Mercury Enrichment and Detection with Flow Cytometry

Pyrrolo-C as a fluorescent probe for monitoring RNA secondary structure formation

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