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Amine-Reactive BODIPY Dye: Spectral Properties and Application for Protein Labeling
Ksenia V Ksenofontova1, Anastasia A Kerner1, Alexander A Ksenofontov1,2
1Department of Inorganic Chemistry, Ivanovo State University of Chemistry and Technology, 10, Sheremetevskiy pr., 153000 Ivanovo, Russia.
Molecules (Basel, Switzerland)
|November 26, 2022
Summary
A novel fluorescent dye, NHS-Ph-BODIPY, was synthesized for protein labeling. This dye exhibits enhanced sensitivity for detecting proteins like BSA, offering a valuable tool for biochemical analysis.
Area of Science:
- Organic Chemistry
- Biochemistry
- Photophysics
Background:
- Boron-dipyrromethene (BODIPY) dyes are widely used fluorescent probes.
- Developing novel fluorescent probes with tailored properties is crucial for biochemical applications.
- Understanding structure-property relationships in fluorescent dyes enhances their utility.
Purpose of the Study:
- To synthesize and characterize a novel amine-reactive BODIPY derivative (NHS-Ph-BODIPY).
- To investigate the spectroscopic and photophysical properties of NHS-Ph-BODIPY and its precursor.
- To evaluate the performance of NHS-Ph-BODIPY in fluorescent labeling and detection of proteins.
Main Methods:
- Synthesis of NHS-Ph-BODIPY and COOH-Ph-BODIPY.
- Spectroscopic and photophysical characterization in various organic solvents.
- Quantum-chemical calculations to understand solvent effects and substituent influence.
- Protein labeling experiments with bovine serum albumin (BSA) and PotN.
- Molecular docking to predict binding sites and energies.
Main Results:
- NHS-Ph-BODIPY and COOH-Ph-BODIPY exhibit green fluorescence with good molar absorption coefficients and quantum yields.
- Solvent polarizability and dipolarity significantly influence the absorption and emission maxima.
- NHS-Ph-BODIPY successfully labels proteins, causing spectral shifts and fluorescence quenching.
- Enhanced sensitivity for BSA detection compared to Coomassie brilliant blue.
- Similar sensitivity for PotN detection with both NHS-Ph-BODIPY and Coomassie brilliant blue.
Conclusions:
- NHS-Ph-BODIPY is a versatile fluorescent probe for protein labeling and detection.
- Solvent effects play a critical role in the photophysical properties of these BODIPY dyes.
- The study provides insights into the binding interactions of NHS-Ph-BODIPY with proteins.
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