Zlotnikov, I.D.; Savchenko, I.V.; Kudryashova, E.V. FRET Probes in Micellar Systems: Micelle Formation Indicator and the Enzymes Catalytic Activity Signal Enhancer. Preprints2023, 2023120409. https://doi.org/10.20944/preprints202312.0409.v1
APA Style
Zlotnikov, I.D., Savchenko, I.V., & Kudryashova, E.V. (2023). FRET Probes in Micellar Systems: Micelle Formation Indicator and the Enzymes Catalytic Activity Signal Enhancer. Preprints. https://doi.org/10.20944/preprints202312.0409.v1
Chicago/Turabian Style
Zlotnikov, I.D., Ivan V. Savchenko and Elena Vadimovna Kudryashova. 2023 "FRET Probes in Micellar Systems: Micelle Formation Indicator and the Enzymes Catalytic Activity Signal Enhancer" Preprints. https://doi.org/10.20944/preprints202312.0409.v1
Abstract
Fluorescent labels, especially FRET probes, are promising tools for studying a number of bio-chemical processes. In this paper, we promoted 2 important applications of FRET probes (MUTMAC-R6G and FITC-R6G): i) the formation of micelles from surfactants of various natures and polymers (chitosan – fatty acid), as well as ii) monitoring of enzymatic activity with improved parameters (increased analytical signal and improved selectivity due to shift to the long-wavelength region). The formation of micelles is accompanied by the convergence of fluorophores in the hydrophobic micelle core by a distance closer than in the buffer solution, thus r/R0 (where R0 – Foerster radius) is chosen as an analytical signal of micelle formation, including critical micelle concentration (CMC) and critical pre-micelle concentration (CPMC). The CMC values calculated using FRET probes are in a good agreement with literature data. At the same time, the r/R0 function provides valuable information about the nature and mechanism of micelle formation. With the second analytical application of FRET probes, we considered the optimization of techniques for studying enzymatic activity. The enzyme catalyses the reaction with the release of a fluorescent product, the signal from which may not be enough for detection, or it may be quenched, for example, in the reverse micelles (AOT-octane). Here we have proposed a solution – a FRET probe containing a rhodamine 6G (R6G) acceptor, which allows us to monitor the enzymatic reaction selectively (in the red region, 550-600 nm), and obtain a significantly higher fluorescence yield (potentially from 10 to 250 times). Thus, we have demonstrated a high potential for the FRET probes application as indicators of micelle formation as well as for the study of the enzyme catalytic activity. In the future, the method developed has prospects for application in the visualization of the enzyme functioning in cells due to the shift of the fluorescence signal to the long-wavelength region with an increase in the signal selectivity to suppress autofluorescence.
Biology and Life Sciences, Biology and Biotechnology
Copyright:
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