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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Engineering the metal sensitive sites in Macrolampis sp2 firefly luciferase and use as a novel bioluminescent ratiometric biosensor for heavy metals

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Author(s):
Gabriel, Gabriele V. M. ; Viviani, Vadim R.
Total Authors: 2
Document type: Journal article
Source: ANALYTICAL AND BIOANALYTICAL CHEMISTRY; v. 408, n. 30, p. 8881-8893, DEC 2016.
Web of Science Citations: 2
Abstract

Most luminescent biosensors for heavy metals are fluorescent and rely on intensity measurements, whereas a few are ratiometric and rely on spectral changes. Bioluminescent biosensors for heavy metals are less common. Firefly luciferases have been coupled to responsive promoters for mercury and arsenium, and used as light on biosensors. Firefly luciferase bioluminescence spectrum is naturally sensitive to heavy metal cations such as zinc and mercury and to pH. Although pH sensitivity of firefly luciferases was shown to be useful for ratiometric estimation of intracellular pH, its potential use for ratiometric estimation of heavy metals was never considered. Using the yellow-emitting Macrolampis sp2 firefly luciferase and site-directed mutagenesis, we show that the residues H310 and E354 constitute two critical sites for metal sensitivity that can be engineered to increase sensitivity to zinc, nickel, and mercury. A linear relationship between cation concentration and the ratio of bioluminescence intensities at 550 and 610 nm allowed, for the first time, the ratiometric estimation of heavy metals concentrations down to 0.10 mM, demonstrating the potential applicability of firefly luciferases as enzymatic and intracellular ratiometric metal biosensors. (AU)

FAPESP's process: 11/23961-0 - pH-sensitive luciferases: structural origin of pH-sensitivity, and suitability of use in cell biosensors for pH and heavy metals
Grantee:Vadim Viviani
Support Opportunities: Regular Research Grants
FAPESP's process: 14/04477-9 - Developing of bioluminescent ratiometric biosensors for pH and divalent metals based on the engineering of the pH-sensing moiety of firefly luciferases
Grantee:Gabriele Verônica de Mello Gabriel
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 13/09594-0 - Analysis of coleopteran lanterns, fat body and Malpighian tubules transcriptomes, and its relationship with the origins of bioluminescence in Coleoptera and Diptera
Grantee:Vadim Viviani
Support Opportunities: Regular Research Grants