Nano-frfast: design of a new genetically-encoded far-red fluorescent label

Мұқаба

Дәйексөз келтіру

Толық мәтін

Ашық рұқсат Ашық рұқсат
Рұқсат жабық Рұқсат берілді
Рұқсат жабық Рұқсат ақылы немесе тек жазылушылар үшін

Аннотация

We propose a new fluorogen-activating protein nano-frFAST with only 98 amino acids based on a combination of the previously created fluorogen-activating proteins nanoFAST and frFAST. A series of fluorogens with an increased system of conjugated bonds, which are potentially capable of binding to this protein, were synthesized. Based on the results of the study, a promising fluorogen which is (Z)-5-((E)-3-(4-hydroxy-2,5-dimethoxyphenyl)allylidene)-2-thioxothiazolidin-4-one (HPAR-DOM) was identified. We demonstrate that the nano-frFAST–HPAR-DOM complex can be used as a genetically encoded far-red fluorescent label for staining individual structures of living cells.

Толық мәтін

Рұқсат жабық

Авторлар туралы

E. Sokolinskaya

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Pirogov Russian National Research Medical University

Хат алмасуға жауапты Автор.
Email: svetlanakr2002@mail.ru
Ресей, Moscow; Moscow

Yu. Bogdanova

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Pirogov Russian National Research Medical University

Email: svetlanakr2002@mail.ru
Ресей, Moscow; Moscow

I. Myasnyanko

Pirogov Russian National Research Medical University

Email: svetlanakr2002@mail.ru
Ресей, Moscow

A. Sokolov

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Pirogov Russian National Research Medical University

Email: svetlanakr2002@mail.ru
Ресей, Moscow; Moscow

S. Krasnova

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS

Email: svetlanakr2002@mail.ru
Ресей, Moscow

M. Baranov

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Pirogov Russian National Research Medical University

Email: svetlanakr2002@mail.ru
Ресей, Moscow; Moscow

Әдебиет тізімі

  1. Snapp E.L. // Trends Cell Biol. 2009. V. 19. P. 649–655. https://doi.org/10.1016/j.tcb.2009.08.002
  2. Chudakov D.M., Matz M.V., Lukyanov S., Lukyanov K.A. // Physiol. Rev. 2010. V. 90. P. 1103–1163. https://doi.org/10.1152/physrev.00038.2009
  3. Hori Y., Ueno H., Mizukami S., Kikuchi K. // J. Am. Chem. Soc. 2009. V. 131. P. 16610–16611. https://doi.org/10.1021/ja904800k
  4. Mineev K.S., Goncharuk S.A., Goncharuk M.V., Povarova N.V., Sokolov A.I., Baleeva N.S., Smirnov A.Yu., Myasnyanko I.N., Ruchkin D.A., Bukhdruker S., Remeeva A., Mishin A., Borshchevskiy V., Gordeliy V., Arseniev A.S., Gorbachev D.A., Gavrikov A.S., Mishin A.S., Baranov M.S. // Chem. Sci. 2021. V. 12. P. 6719–6725. https://doi.org/10.1039/d1sc01454d
  5. Plamont M.-A., Billon-Denis E., Maurin S., Gauron C., Pimenta F.M., Specht C.G., Shi J., Quérard J., Pan B., Rossignol J., Moncoq K., Morellet N., Volovitch M., Lescop E., Chen Y., Triller A., Vriz S., Saux T. Le, Jullien L., Gautier A. // Proc. Natl. Acad. Sci. USA. 2016. V. 113. P. 497–502. https://doi.org/10.1073/pnas.1513094113
  6. Hocq R., Bottone S., Gautier A., Pflügl S. // Front. Bioeng. Biotechnol. 2023. V. 11. P. 1226889. https://doi.org/10.3389/fbioe.2023.1226889
  7. Tsien R.Y. // Annu. Rev. Biochem. 1998. V. 67. P. 509–544. https://doi.org/10.1146/annurev.biochem.67.1.509
  8. Remington S.J. // Curr. Opin. Struct. Biol. 2006. V. 16. P. 714–721. https://doi.org/10.1016/j.sbi.2006.10.001
  9. Benaissa H., Ounoughi K., Aujard I., Fischer E., Goïame R., Nguyen J., Tebo A.G., Li C., Le Saux T., Bertolin G., Tramier M., Danglot L., Pietrancosta N., Morin X., Jullien L., Gautier A. // Nat. Commun. 2021. V. 12. P. 6989. https://doi.org/10.1038/s41467-021-27334-0
  10. Myasnyanko I.N., Gavrikov A.S., Zaitseva S.O., Smirnov A.Yu., Zaitseva E.R., Sokolov A.I., Malyshevskaya K.K., Baleeva N.S., Mishin A.S., Baranov M.S. // Chemistry. 2021. V. 27. P. 3986–3990. https://doi.org/10.1002/chem.202004760
  11. Povarova N.V., Zaitseva S.O., Baleeva N.S., Smirnov A.Yu., Myasnyanko I.N., Zagudaylova M.B., Bozhanova N.G., Gorbachev D.A., Malyshevskaya K.K., Gavrikov A.S., Mishin A.S., Baranov M.S. // Chemistry. 2019. V. 25. P. 9592–9596. https://doi.org/10.1002/chem.201901151
  12. Chen C., Tachibana S.R., Baleeva N.S., Myasnyanko I.N., Bogdanov A.M., Gavrikov A.S., Mishin A.S., Malyshevskaya K.K., Baranov M.S., Fang C. // Chemistry. 2021. V. 27. P. 8946–8950. https://doi.org/10.1002/chem.202101250
  13. Li C., Tebo A.G., Thauvin M., Plamont M.-A., Volovitch M., Morin X., Vriz S., Gautier A. // Angew. Chem. Int. Ed. Engl. 2020. V. 59. P. 17917–17923. https://doi.org/10.1002/anie.202006576
  14. Baleeva N.S., Bogdanova Y.A., Goncharuk M.V., Sokolov A.I., Myasnyanko I.N., Kublitski V.S., Smirnov A.Yu., Gilvanov A.R., Goncharuk S.A., Mineev K.S., Baranov M.S. // Int. J. Mol. Sci. 2024. V. 25. Р. 3054. https://doi.org/10.3390/ijms25053054
  15. Perfilov M.M., Zaitseva E.R., Smirnov A.Yu., Mikhaylov A.A., Baleeva N.S., Myasnyanko I.N., Mishin A.S., Baranov M.S. // Dyes Pigm. 2022. V. 198. P. 110033. https://doi.org/10.1016/j.dyepig.2021.110033
  16. Campaigne E., White R.L. // J. Heterocycl. Chem. 1988. V. 25. P. 367–373. https://doi.org/10.1002/jhet.5570250203
  17. Radi M., Botta L., Casaluce G., Bernardini M., Botta M. // J. Comb. Chem. 2010. V. 12. P. 200–205. https://doi.org/10.1021/cc9001789
  18. Voliani V., Bizzarri R., Nifosì R., Abbruzzetti S., Grandi E., Viappiani C., Beltram F. // J. Phys. Chem. B. 2008. V. 112. P. 10714–10722. https://doi.org/10.1021/jp802419h
  19. Würth C., Grabolle M., Pauli J., Spieles M., ReschGenger U. // Nat. Protoc. 2013. V. 8. P. 1535–1550. https://doi.org/10.1038/nprot.2013.087

Қосымша файлдар

Қосымша файлдар
Әрекет
1. JATS XML
Жүктеу
2. Additional materials
Жүктеу (2MB)
Метадеректер
3. Fig. 1. Absorption and emission spectra of the nano-frFAST–HPAR-DOM complex in phosphate buffer.

Жүктеу (188KB)
Метадеректер
4. Fig. 2. Representative photomicrographs of Hela Kyoto cells expressing the H2B-nano-frFAST (upper panel) and hCytokeratin-nano-frFAST (lower panel) genetic constructs in the presence of 10 μM HPAR-DOM fluorogen. Scale bar is 10 μm.

Жүктеу (207KB)
Метадеректер
5. Scheme 1. Structural formulas of known fluorogens of the FAST and frFAST proteins, a new proposed fluorogen for the developed nano-frFAST protein, as well as approaches to synthesis.

Жүктеу (313KB)
Метадеректер

© Russian Academy of Sciences, 2025