Journal of Radioanalytical and Nuclear Chemistry

, Volume 286, Issue 3, pp 835–840 | Cite as

In vitro study of proteins surface activity by tritium probe

  • Maria G. Chernysheva
  • Gennadii A. Badun


A new technique for in vitro studies of biomacromolecules interactions, their adsorption at aqueous/organic liquid interfaces and distribution in the bulk of liquid/liquid systems was developed. The method includes (1) tritium labeling of biomolecules by tritium thermal activation method and (2) scintillation phase step with organic phase, which can be concerned as a model of cellular membrane. Two globular proteins lysozyme and human serum albumin tested. We have determined the conditions of tritium labeling when labeled by-products can be easy separated by means of dialysis and size-exclusion chromatography. Scintillation phase experiments were conducted for three types of organic liquids. Thus, the influences of the nature of organic phase on proteins adsorption and its distribution in the bulk of aqueous/organic liquid system were determined. It was found that proteins possess high surface activity at aqueous/organic liquid interface. Furthermore, values of hydrophobicity of globular proteins were found by the experiment.


Tritium labeled macromolecules Scintillation phase method 



This work was supported by Russian Foundation of Basic Researches (grant # 09-03-00819) and by Federal Targeted Program “Scientific and scientific-pedagogical personnel of innovation Russia” for 2009–2013 years (project # 2351P).


  1. 1.
    Růžiška M, Borecký J, Hanuš J, Ježek P (1996) FEBS Lett 382:239CrossRefGoogle Scholar
  2. 2.
    Xie H, Audette C, Hoffee M, Lambert JM, Blättler WA (2004) J Pharmacol Exp Ther 308:1073CrossRefGoogle Scholar
  3. 3.
    Sengupta T, Damodaran S (1998) J Colloids Interface Sci 206:407CrossRefGoogle Scholar
  4. 4.
    Graham DE, Phillips MC (1979) J Colloid Interface Sci 70:403CrossRefGoogle Scholar
  5. 5.
    Graham DE, Phillips MC (1979) J Colloid Interface Sci 70:414Google Scholar
  6. 6.
    Scott Wilbur D (1992) Bioconjug Chem 3:433CrossRefGoogle Scholar
  7. 7.
    Sengupta T, Razumovsky L, Damodaran S (1999) Langmuir 15:6991CrossRefGoogle Scholar
  8. 8.
    Sengupta T, Damodaran S (1998) Langmuir 14:6457CrossRefGoogle Scholar
  9. 9.
    Damodaran S, Sengupta T (2003) J Agric Food Chem 51:1658CrossRefGoogle Scholar
  10. 10.
    Feinendegen LE (1967) Tritium-labeled molecules in biology and chemistry. Academic Press, New YorkGoogle Scholar
  11. 11.
    Steinberg D, Vaughan M, Anfinsen CB, Gorry JD (1957) Science 126:447CrossRefGoogle Scholar
  12. 12.
    Tack BF, Dean J, Eilat D, Lorem PE, Schechter AN (1980) J Biol Chem 255:8842Google Scholar
  13. 13.
    Baratova LA, Efimov AV, Dobrov EN, Fedorova NV, Hunt R, Badun GA, Ksenofontov AL, Torrance L, Järvekülg L (2001) J Virol 75:9696CrossRefGoogle Scholar
  14. 14.
    Shishkov AV, Goldanskii VI, Baratova LA, Fedorova NV, Ksenofontov AL, Zhirnov OP, Galkin AV (1999) Proc Natl Acad Sci USA 96:7827CrossRefGoogle Scholar
  15. 15.
    Dobrov EN, Badun GA, Lukashina EV, Fedorova NV, Ksenofontov AL, Fedoseev VM, Baratova LA (2003) Eur J Biochem 270:3300CrossRefGoogle Scholar
  16. 16.
    Lukashina E, Badun G, Fedorova N, Ksenofontov A, Nemykh M, Serebryakova M, Mukhamedzhanova A, Karpova O, Rodionova N, Baratova L, Dobrov E (2009) FEBS J 276:7006CrossRefGoogle Scholar
  17. 17.
    Lukashina EV, Badun GA, Chulichkov AL (2007) Biomol Eng 24:125CrossRefGoogle Scholar
  18. 18.
    Chernysheva MG, Tyasto ZA, Badun GA (2009) J Radioanal Nucl Chem 280:303CrossRefGoogle Scholar
  19. 19.
    Badun GA, Chernysheva MG, Pozdnyakova VYu, Fedoseev VM (2005) Radiochemistry 47:584CrossRefGoogle Scholar
  20. 20.
    Badun GA, Chernysheva MG, Tyasto ZA, Fedoseev VM (2009) Mosc Univ Chem Bull 64:282CrossRefGoogle Scholar
  21. 21.
    Badun GA, Chernysheva MG, Tyasto ZA, Kulikova NA, Kudryavtsev AV, Perminova IV (2010) Radiochim Acta 98:161CrossRefGoogle Scholar
  22. 22.
    Chernysheva MG, Badun GA (2009) Proceedings of the VIII Finnish-Russian symposium on radiochemistry, Turku, Finland, 4–5 Sept 2009, p 94Google Scholar
  23. 23.
    Badun GA, Chernysheva MG (2009) Proceedings of the VIII Finnish-Russian symposium on radiochemistry, Turku, Finland, 4–5 Sept 2009, p 96Google Scholar
  24. 24.
    Shishkov AV, Neiman LA, Smolyakov VS (1984) Russ Chem Rev 53:656CrossRefGoogle Scholar
  25. 25.
    Chernysheva MG, Badun GA, Tyasto ZA, Pozdnyakova VYu, Fedoseev VM, Ksenofontov AL (2007) Radiochemistry 49:186CrossRefGoogle Scholar
  26. 26.
    Bigelow CC (1967) J Theor Biol 16:187CrossRefGoogle Scholar

Copyright information

© Akadémiai Kiadó, Budapest, Hungary 2010

Authors and Affiliations

  1. 1.Chemistry Department, Radiochemistry DivisionLomonosov Moscow State UniversityMoscowRussia

Personalised recommendations