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Measuring nanoparticle-induced resonance energy transfer effect by electrogenerated chemiluminescent reactions.

ABSTRACT: Electrogenerated chemiluminescence (ECL) efficiencies, redox potentials, photoluminescent (PL) (quenching and coupling) effects, and AFM images for the [Ru(bpy)₃]²⁺/Au@tiopronin system were determined in aqueous solutions of the gold nanoparticles (NPs) at pH 7.0. The most remarkable finding was that ECL measurements can display the nanoparticle-induced resonance energy transfer (NP-RET) effect. Its effectiveness was quantified through a coefficient, K _{(NP-RET)_ECL} , which measures how much an ECL reaction has been enhanced. Moreover, the NP-RET effect was also checked using PL measurements, in such a way that a coefficient, K _{(NP-RET)_PL} , was determined; both constants, K _{(NP-RET)_ECL} and K _{(NP-RET)_PL} being in close agreement. It is important to highlight the fact that the NP-RET effect is only displayed in diluted solutions in which there is no NPs self-aggregation. The existence of the NPs self-aggregation behavior is revealed through AFM measurements.

SUBMITTER: Perez-Tejeda P

PROVIDER: S-EPMC9048710 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Measuring nanoparticle-induced resonance energy transfer effect by electrogenerated chemiluminescent reactions.

Perez-Tejeda Pilar P Martínez-Delgado Alberto A Grueso Elia E Giráldez-Pérez Rosa M RM

RSC advances 20200123 7

Electrogenerated chemiluminescence (ECL) efficiencies, redox potentials, photoluminescent (PL) (quenching and coupling) effects, and AFM images for the [Ru(bpy)3]2+/Au@tiopronin system were determined in aqueous solutions of the gold nanoparticles (NPs) at pH 7.0. The most remarkable finding was that ECL measurements can display the nanoparticle-induced resonance energy transfer (NP-RET) effect. Its effectiveness was quantified through a coefficient, K (NP-RET)< ...[more]

PMID: 35492653

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Project description:UnlabelledProtein-protein interactions are important for virtually every biological process, and a number of elegant approaches have been designed to detect and evaluate such interactions. However, few of these methods allow the detection of dynamic and real-time protein-protein interactions in bacteria. Here we describe a bioluminescence resonance energy transfer (BRET) system based on the bacterial luciferase LuxAB. We found that enhanced yellow fluorescent protein (eYFP) accepts the emission from LuxAB and emits yellow fluorescence. Importantly, BRET occurred when LuxAB and eYFP were fused, respectively, to the interacting protein pair FlgM and FliA. Furthermore, we observed sirolimus (i.e., rapamycin)-inducible interactions between FRB and FKBP12 and a dose-dependent abolishment of such interactions by FK506, the ligand of FKBP12. Using this system, we showed that osmotic stress or low pH efficiently induced multimerization of the regulatory protein OmpR and that the multimerization induced by low pH can be reversed by a neutralizing agent, further indicating the usefulness of this system in the measurement of dynamic interactions. This method can be adapted to analyze dynamic protein-protein interactions and the importance of such interactions in bacterial processes such as development and pathogenicity.ImportanceReal-time measurement of protein-protein interactions in prokaryotes is highly desirable for determining the roles of protein complex in the development or virulence of bacteria, but methods that allow such measurement are not available. Here we describe the development of a bioluminescence resonance energy transfer (BRET) technology that meets this need. The use of endogenous excitation light in this strategy circumvents the requirement for the sophisticated instrument demanded by standard fluorescence resonance energy transfer (FRET). Furthermore, because the LuxAB substrate decanal is membrane permeable, the assay can be performed without lysing the bacterial cells, thus allowing the detection of protein-protein interactions in live bacterial cells. This BRET system added another useful tool to address important questions in microbiological studies.

Dataset Information

Measuring nanoparticle-induced resonance energy transfer effect by electrogenerated chemiluminescent reactions.

Publications

Measuring nanoparticle-induced resonance energy transfer effect by electrogenerated chemiluminescent reactions.

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