Selectivity and Sensitivity of Fluorescence Lifetime-Based Metal Ion Biosensing Using a Carbonic Anhydrase Transducer

doi:10.1006/abio.1998.2991

Analytical Biochemistry

Volume 267, Issue 1, 1 February 1999, Pages 185-195

https://doi.org/10.1006/abio.1998.2991 Get rights and content

Abstract

A key performance criterion for metal ion determinations in complex media like serum, cytoplasm of the cell, and sea water is selectivity: the ability to determine the analyte(s) of interest, in the presence of relatively high concentrations of interferents. Cu(II), Zn(II), Cd(II), Co(II), and Ni(II) may be determined by changes they induce in the fluorescence lifetime and intensity of site-specifically labeled fluorescent variants of apocarbonic anhydrase II. Free metal ion concentrations in the picomolar range (for Cu(II) and Zn(II)) and the nanomolar range (for Cd(II), Co(II), and Ni(II)) were determined, based on the affinity of the apoenzyme for these ions. Mg(II) at 50 mM and Ca(II) at 10 mM produced no effect. By the use of different fluorescent labels, transducers were made which responded well to Cu(II), Co(II), and Ni(II), but not to Zn(II) and Cd(II), and vice versa.

References (41)

M.P. Cuajungco et al.
Neurobiol. Disc.
(1997)
R.B. Thompson et al.
Biosens. Bioelectr.
(1996)
S. Lindskog et al.
Biochim. Biophys. Acta
(1964)
T.J.B. Simons
J. Biochem. Biophys. Methods
(1993)
G. Grynkiewicz et al.
J. Biol. Chem.
(1985)
R.B. Thompson et al.
Anal. Biochem.
(1995)
J.E. Coleman
J. Biol. Chem.
(1967)
S.K. Nair et al.
J. Biol. Chem.
(1991)
S.K. Nair et al.
J. Biol. Chem.
(1996)
R.F. Chen et al.
J. Biol. Chem.
(1967)

C.J. Frederickson

Int. Rev. Neurobiol.

(1989)

R.C. Prince et al.

Trends Biochem. Sci.

(1998)

J.A. Hunt et al.

J. Biol. Chem.

(1997)

T.V. O'Halloran

Science

(1993)

D.W. Choi et al.

Annu. Rev. Neurosci.

(1998)

R.Y. Tsien

Annual Review of Neuroscience

(1989)

S. Lindskog et al.

The Enzymes

(1971)

R.B. Thompson et al.

Anal. Chem.

(1993)

R.B. Thompson

R.B. Thompson et al.

J. Fluoresc.

(1995)

Cited by (35)

Picomolar level sensorial dual colorimetric gold nanoparticle sensor for Zn<sup>2+</sup> and Hg<sup>2+</sup> ions synthesized from bark extract of Lannea Grandis Coromandelica and its wide range applications in real sample analysis
2024, Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy
In this work a facile, rapid, reproducible and non-toxic approach has been demonstrated for synthesis of most stable AuNPs from bark extract of Lannea Grandis Coromandelica. UV–Visible spectroscopy, FTIR, TEM, SAED, EDX, XRD, DLS, Zeta Potential, FE-SEM, AFM and XPS techniques were employed for the characterization of synthesized LGC-AuNPs. The UV–Vis spectra of LGC-AuNPs gave SPR peak at 536 nm while the TEM analysis revealed LGC-AuNPs have 20.75 nm size with spherical in shape. DLS study showed the AuNPs have average diameter 50.18 nm. The synthesized AuNPs exhibited very high selectivity, rapid response in recognition towards Zn²⁺ and Hg²⁺ ions by changing its color within 20 sec. This proposed sensor can detect very low picomolar level of Zn²⁺ and Hg²⁺ ions (LOD value for Zn²⁺ and Hg²⁺ were found 1.36 pM and 24.60 pM respectively). Here we also studied effect of several factors such as variation of conc of gold, temperature, incubation time, pH, salt, solvent (polar protic and polar aprotic) to know in which condition AuNPs have high stability and sensitivity. The data revealed that synthesized AuNPs was stable up to two years at pH 6.5 at room temperature in water media and under this condition, it shows maximum sensitivity and reactivity. Moreover, here interference study was carried out to identify high selectivity of synthesized LGC-AuNPs probe in presence of different metal ions. The real sample analyses also revealed the great applicability of this probe. Therefore, this simple, rapid, low-cost, sensing activity appeared to hold great sensibleness for detection of heavy metal ions in real sample.
Analytical study on cofactor biorecognition by immobilized alkaline apophosphatase
2011, Sensors and Actuators, B: Chemical
Citation Excerpt :
ApoCA-Zn biosensing scheme under FIA conditions with multiple use of immobilized enzyme has been used for environmental analysis [12]. A number of studies on highly selective and ultrasensitive fluorimetric detection of zinc ions with apoCA have been shown and reviewed only recently [13–15]. Zinc biodetermination with apoenzyme receptors is also possible using aminopeptidase [16], carboxypeptidase [17], thermolysin [18] and alcohol dehydrogenases [19].
Alkaline apophosphatase (apoALP) was examined in flow bioanalytical systems as an immobilized bioligand for reversible recognition of zinc ions. For such investigations plastic open-tubular bioreactor coupled with spectrophotometric detector and potentiometric biosensor based on plasticized polymeric membrane ion-selective electrode were applied. Both biodevices contain covalently immobilized monomolecular layer of alkaline phosphatase (ALP) and operate according to the indirect biosensing scheme based on cofactor biorecognition by in situ generated apoenzymatic bioreceptor. An implementation of immobilized ALP into flow system enables automation of the developed bioanalytical procedure allowing multiple and reproducible apoenzyme formation/regeneration, analyte biorecognition as well as assays for recovered enzyme activity. Both developed bioanalytical systems offer relatively fast zinc biodetermination in the ppm range of concentrations. Additionally, effects from potential chelators and metal ions involving in holo–apo and apo–holo enzyme conversions were investigated. It was found, that the selectivity of the reported biosensing scheme is defined by alternative mechanisms for apoenzyme and holoenzyme formation. A model, clarifying such conversions, has been proposed and confirmed by simple analytical experiments performed under flow conditions.
Carbonic anhydrase II-based metal ion sensing: Advances and new perspectives
2010, Biochimica et Biophysica Acta - Proteins and Proteomics
Carbonic anhydrases are archetypical zinc metalloenzymes and as such, they have been developed as the recognition element of a family of fluorescent indicators (sensors) to detect metal ions, particularly Zn²⁺ and Cu²⁺. Subtle modification of the structure of human carbonic anhydrase II isozyme (CAII) alters the selectivity, sensitivity, and response time for these sensors. Sensors using CAII variants coupled with zinc-dependent fluorescent ligands demonstrate picomolar sensitivity, unmatched selectivity, ratiometric fluorescence signal, and near diffusion-controlled response times. Recently, these sensors have been applied to measuring the readily exchangeable concentrations of zinc in the cytosol and nucleus of mammalian tissue culture cells and concentrations of free Cu²⁺ in seawater.
Studying zinc biology with fluorescence: Ain't we got fun?
2005, Current Opinion in Chemical Biology
Zinc has emerged as a metal ion of substantial interest in biology and medicine, especially in neuroscience, gene transcription, the immune response, and mammalian reproduction. Fueling these advances in understanding has been the development of new fluorescence-based indicator systems for zinc with unprecedented sensitivity and selectivity. This review summarizes recent progress in the development of fluorescence-based sensors and biosensors for zinc, with a view to evaluating their suitability for use with biologically derived specimens, especially in vivo and in situ.
Divalent metal ion-sensitive holographic sensors
2005, Analytica Chimica Acta
Citation Excerpt :
However, whilst all these technologies are highly sensitive, they are time-consuming and require expensive instrumentation and large sample volumes. In the last few years, there has been an increased interest in circumventing these problems by developing new analytical methods [5–17] and fabricating mass-producible, inexpensive and small sample volume sensors for real-time monitoring of divalent metal ions. Optical chemosensors have been developed for the detection of metal ions in which changes in absorbance or fluorescent properties of different chelating compounds are determined when binding the metal.
A holographic sensor for real-time detection of divalent metal ions (Ca²⁺, Mg²⁺, Ni²⁺, Co²⁺ and Zn²⁺) has been fabricated by incorporating a chelating monomer into a hydrogel matrix. A methacrylated analogue of iminodiacetic acid (IDA) was prepared and co-polymerised with hydroxyethyl methacrylate (HEMA) and ethylene glycol dimethacrylate (EDMA) as a cross-linker to form polymer films. A silver-based reflection hologram was incorporated into the hydrogel by diffusion followed by holographic recording using a frequency-doubled Nd/YAG laser. Changes in the replay wavelength of the hologram were used to characterise the swelling behaviour of the matrix as a function of its chemical composition and concentration of analyte in the media. The effects of active monomer, cross-linker, pH and ionic strength on the swelling of the matrix and on metal detection sensitivity have been studied. Polymers containing >10 mol% of chelating monomer and 6 mol% of cross-linker showed significant responses (46.3 nm) within 30 s at an ion concentration of 0–40 nm. The selectivity of the holograms towards the different ions tested was Ni²⁺>Zn²⁺>Co²⁺>Ca²⁺>Mg²⁺. The sensor showed fully reversible responses, permitting real-time monitoring of calcium ion efflux during the germination of Bacillus megaterium spores.
Chapter 10 Non-affinity sensing technology: the exploitation of biocatalytic events for environmental analysis
2005, Comprehensive Analytical Chemistry
This chapter focuses on well-recognized and emerging biocatalytic approaches that can be applied to the screening or monitoring of environmentally relevant chemical species for the assessment of good ecological and chemical status. The chapter discusses classic catalytic approaches and well-characterized enzyme reactions as well as other catalytic events in living systems that elicit useful bioanalytical information through catabolic activities. The exploitation of affinity interactions—in particular, immunoreactions—for environmental applications is presented. The chapter describes biosensors within the environmental area for the selection of specific parameters as well as the evaluation of the new monitoring or screening challenges to preserve the environment. The bioengineering tools that are currently expanding and improving the biocatalytic approach in environmental applications are presented in the chapter.

View all citing articles on Scopus

^☆: Boyer, P. D.

¹: To whom correspondence should be addressed. Fax: (410) 708-7122. E-mail:[email protected]

View full text

Regular ArticleSelectivity and Sensitivity of Fluorescence Lifetime-Based Metal Ion Biosensing Using a Carbonic Anhydrase Transducer☆

Abstract

Neurobiol. Disc.

Biosens. Bioelectr.

Biochim. Biophys. Acta

J. Biochem. Biophys. Methods

J. Biol. Chem.

Anal. Biochem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Int. Rev. Neurobiol.

Trends Biochem. Sci.

J. Biol. Chem.

Science

Annu. Rev. Neurosci.

Annual Review of Neuroscience

The Enzymes

Anal. Chem.

J. Fluoresc.

Regular Article
Selectivity and Sensitivity of Fluorescence Lifetime-Based Metal Ion Biosensing Using a Carbonic Anhydrase Transducer☆