Amine-Reactive BODIPY Dye: Spectral Properties and Application for Protein Labeling
Ksenia V. Ksenofontova,
Anastasia A. Kerner,
Alexander A. Ksenofontov,
Artyom Yu. Shagurin,
Pavel S. Bocharov,
Michael M. Lukanov,
Airat R. Kayumov,
Darya E. Zhuravleva,
Zalina I. Iskhakova,
Evgeniy E. Molchanov,
Dmitriy A. Merkushev,
Ilya A. Khodov,
Yuriy S. Marfin
Affiliations
Ksenia V. Ksenofontova
Department of Inorganic Chemistry, Ivanovo State University of Chemistry and Technology, 10, Sheremetevskiy pr., 153000 Ivanovo, Russia
Anastasia A. Kerner
Department of Inorganic Chemistry, Ivanovo State University of Chemistry and Technology, 10, Sheremetevskiy pr., 153000 Ivanovo, Russia
Alexander A. Ksenofontov
Department of Inorganic Chemistry, Ivanovo State University of Chemistry and Technology, 10, Sheremetevskiy pr., 153000 Ivanovo, Russia
Artyom Yu. Shagurin
Laboratoire de Spectroscopie pour les Interactions, la Réactivité et l’Environnement (UMR CNRS 8516), Université de Lille, CEDEX, 59655 Villeneuve d’Ascq, France
Pavel S. Bocharov
G. A. Krestov Institute of Solution Chemistry of The Russian Academy of Sciences, 1, ul. Akademicheskaya, 153045 Ivanovo, Russia
Michael M. Lukanov
Department of Inorganic Chemistry, Ivanovo State University of Chemistry and Technology, 10, Sheremetevskiy pr., 153000 Ivanovo, Russia
Airat R. Kayumov
Institute of Fundamental Medicine and Biology, Kazan Federal University, 18, ul. Kremlyovskaya, 420008 Kazan, Russia
Darya E. Zhuravleva
Institute of Fundamental Medicine and Biology, Kazan Federal University, 18, ul. Kremlyovskaya, 420008 Kazan, Russia
Zalina I. Iskhakova
Institute of Fundamental Medicine and Biology, Kazan Federal University, 18, ul. Kremlyovskaya, 420008 Kazan, Russia
Evgeniy E. Molchanov
Department of Fine Organic Synthesis Technology, Ivanovo State University of Chemistry and Technology, 7, Sheremetevskiy pr., 153000 Ivanovo, Russia
Dmitriy A. Merkushev
Department of Inorganic Chemistry, Ivanovo State University of Chemistry and Technology, 10, Sheremetevskiy pr., 153000 Ivanovo, Russia
Ilya A. Khodov
G. A. Krestov Institute of Solution Chemistry of The Russian Academy of Sciences, 1, ul. Akademicheskaya, 153045 Ivanovo, Russia
Yuriy S. Marfin
Department of Inorganic Chemistry, Ivanovo State University of Chemistry and Technology, 10, Sheremetevskiy pr., 153000 Ivanovo, Russia
A boron-dipyrromethene (BODIPY) derivative reactive towards amino groups of proteins (NHS-Ph-BODIPY) was synthesized. Spectroscopic and photophysical properties of amine-reactive NHS-Ph-BODIPY and its non-reactive precursor (COOH-Ph-BODIPY) in a number of organic solvents were investigated. Both fluorescent dyes were characterized by green absorption (521–532 nm) and fluorescence (538–552 nm) and medium molar absorption coefficients (46,500–118,500 M−1·cm−1) and fluorescence quantum yields (0.32 – 0.73). Solvent polarizability and dipolarity were found to play a crucial role in solvent effects on COOH-Ph-BODIPY and NHS-Ph-BODIPY absorption and emission bands maxima. Quantum-chemical calculations were used to show why solvent polarizability and dipolarity are important as well as to understand how the nature of the substituent affects spectroscopic properties of the fluorescent dyes. NHS-Ph-BODIPY was used for fluorescent labeling of a number of proteins. Conjugation of NHS-Ph-BODIPY with bovine serum albumin (BSA) resulted in bathochromic shifts of absorption and emission bands and noticeable fluorescence quenching (about 1.5 times). It was demonstrated that the sensitivity of BSA detection with NHS-Ph-BODIPY was up to eight times higher than with Coomassie brilliant blue while the sensitivity of PII-like protein PotN (PotN) detection with NHS-Ph-BODIPY and Coomassie brilliant blue was almost the same. On the basis of the molecular docking results, the most probable binding sites of NHS-Ph-BODIPY in BSA and PotN and the corresponding binding free energies were estimated.
