Resonance energy transfer study of hemoglobin binding to model lipid membranes

  • O. K. Kutsenko V.N. Karazin Kharkov National University
  • G. P. Gorbenko V.N. Karazin Kharkov National University
  • V. M. Trusova V.N. Karazin Kharkov National University
Keywords: hemoglobin, model membranes, protein-lipid complexes, fluorescence energy transfer

Abstract

In the present study fluorescence resonance, energy transfer (FRET) technique was employed to obtain information about the structure of hemoglobin (Hb) complexes with model lipid membranes of different
composition. For this purpose three membrane probes, 3-methoxybenzanthrone (MBA), 4-
dimethylaminochalcone (DMC) and 6-propionyl-2-dimethylaminonaphthalene (Prodan) were assessed as
possible donors for heme moiety of the protein. Model membranes were composed of zwitterionic lipid
phosphatidylcholine (PC), anionic lipid cardiolipin (CL), and cholesterol (Chol). FRET measurements
were interpreted in terms of the model of energy transfer in two-dimensional systems proposed by Fung
and Stryer and further extended by Davenport et al. No FRET was observed between Prodan and Hb
because Prodan under the employed experimental conditions was not distributed into the lipid bilayer. In
the case of DMC, Hb-induced oxidative processes in the lipid phase hampered the estimation of Hb
location in a lipid bilayer. Therefore, structural analysis of Hb-lipid complexes was carried out using
MBA as a donor. First, the donor quantum yield, Förster radii, and fluorescence anisotropy of the probes
have been measured. Second, the amount of Hb bound to model membranes was estimated in terms of the
lattice models of large ligand adsorption to lipid bilayers allowing for the possibility of protein insertion
into the membrane interior. Finally, the distance from the acceptor plane to the bilayer center and the depth of Hb penetration into the lipid bilayer were calculated. It was assumed that protein binds to membranes in the form of dimers and penetrates into the membrane interior. In neutral liposomes, Hb penetrates only to the depth of lipid headgroups. The observed higher extent of Hb penetration into Chol containing bilayer as
compared to PC liposomes may be a consequence of specific Hb-Chol interaction. In the case of PC/CL
liposomes, Hb was found to insert in the non-polar membrane region. Taking into account the possibility
of forming the inverted hexagonal structures in the presence of CL, it cannot be excluded that Hb being
entrapped in such structures, translocates through the membrane. If this phenomenon takes place, deeper
Hb penetration into lipid bilayer might be expected. The obtained results can be useful for exact
characterization of Hb binding to the membranes.

Downloads

Download data is not yet available.

Author Biographies

O. K. Kutsenko, V.N. Karazin Kharkov National University

4 Svobody Sq., Kharkiv, 61077

G. P. Gorbenko, V.N. Karazin Kharkov National University

4 Svobody Sq., Kharkiv, 61077

V. M. Trusova, V.N. Karazin Kharkov National University

4 Svobody Sq., Kharkiv, 61077

References

1. Lakowicz J.R. Principles of fluorescent spectroscopy, Springer: Singapore, 2006, 954 p.

2. Selvin P.R. The renaissance of fluorescence resonance energy transfer // Nat. Struct. Biol. – 2000. – V. 7, N 9. – P. 730-734.

3. Szollosi J., Damjanovich S., Matyus L. Application of fluorescence resonance energy transfer in the clinical laboratory: routine and research // Cytometry – 1998. – V. 34. – P. 159-179.

4. Demehin A.A., Abugo O.O., Jayakumar R., Lakowicz J.R., Rifkind J.M. Binding of hemoglobin to red cell membranes with Eosin-5-maleimide-labeled Band 3: Analysis of centrifugation and fluorescence lifetime data // Biochemistry. – 2002. – V. 41. – P. 8630-8637.

5. Chang T.M.S. Future generation of red blood cell substitutes // J. Internal Medicine. – 2003. – V. 253. – P. 527-535.

6. Fan C., Zhong J., Guan R., Li G. Direct electrochemical characterization of vitreoscilla sp. Hemoglobin entrapped in organic films // Biochim. Biophys. Acta. – 2003. – V. 1649. – P.123-126.

7. Eisinger J., Flores J., Bookchin R.M. The cytosol-membrane interface of normal and sickle erythrocytes // J. Biol. Chem. – 1984. – V. 259, N. 11. – P. 7169-7177.

8. Eisinger J., Flores J. The relative locations of intramembrane fluorescent probes and of the cytosol haemoglobin in erythrocytes, studied by transverse resonance energy transfer // Biophys. J. – 1982. – V. 37. – P. 6-7.

9. Gorbenko G.P. Resonance energy transfer study of haemoglobin complexes with model phospholipid membranes // Biophys. Chem. – 1999. – V. 81. – P. 93-105.

10. Mui B., Chow L., Hope M.J. Extrusion technique to generate liposomes of defined size. // Meth Enzymol. – 2003. – V. 367. – P. 3–14.

11. Dobretsov G.E., Fluorescent probes in the studies of cells, membranes and lipoproteins, Nauka: Moscow, 1989, 277 p.

12. Fung B.K., Stryer L. Surface density determination in membranes by fluorescence energy transfer. // Biochemistry. – 1978. – V. 17. – P. 5241-5248.

13. Davenport L., Dale R., Bisby R., Cundall R. Transverse location of the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene in model lipid bilayer membrane systems by resonance excitation energy transfer // Biochemistry. – 1985. – V. 24. – P. 4097–4108.

14. Dobretsov G.E., Dmitriyev V.M., Pirogova L.B., Petrov V.A., Vladimirov Yu.A. 4-dimethylaminochalcone and 3-methoxybenzantrone as fluorescent probes to study biomembranes. I. Spectral characteristics // Stud. Biophys. – 1977. – V. 65, N. 2. – P. 91-98.

15. Stankowski S. Large-ligand adsorption membranes. III. Cooperativity and general ligand shapes // Biochim. Biophys. Acta. – 1984. – V. 777. – P. 167-182.

16. Shviro Y., Zibber I., Shaklai N. The interaction of hemoglobin with phosphatidylserine vesicles // Biochim. Biophys. Acta. – 1982. – V. 687. – P. 63-70.

17. Bossi L., Alema S., Calissano P., Marra R. Interaction of different forms of hemoglobin with artificial lipid membranes // Biochim. Biophys. Acta. – 1975. – V.375. – P.477-482.

18. Szebeni J., Hauser H., Eskelson C.D., Watson R.R., Winterhalter K.H. Interaction of hemoglobin derivatives with liposomes. Membrane cholesterol protects against the changes of hemoglobin // Biochem. J. – 1988. – V. 27. – P. 6425-6434.

19. Datta P., Chakrabarty S., Chakrabarty A., Chakrabarti A. Membrane interactions of hemoglobin variants, HbA, HbE, HbF and globin subunits of HbA: Effects of aminophospholipids and cholesterol // Biochim. Biophys. Acta. – 2008. – V. 1778. – P. 1-9.

20. LaBrake C.C., Fun L.W.-M. Phospholipid vesicles promote human hemoglobin oxidation // J. Biol. Chem. – 1992. – V. 267, N. 23. – P. 16703-16711.

21. Xi J., Guo R., Guo X. Interactions of hemoglobin with lecithin liposomes // Colloid. Polym. Sci. – 2006. – V. 284. – P. 1139-1145.

22. Guilio A.D., Bonamore A. Globin interactions with lipids and membranes // Methods Enzymol. – 2008. – V. 436. – P. 239-253.

23. Nikolic M., Vranic D., Spiric A., Batas V., Nikolic-Kokic A., Radetic P., Turubatovic L., Blagojevic D.P., Jones D.R., Niketic V., Spasic M.B. Could cholesterol bound to haemoglobin be a missing link for the occasional inverse relationship between superoxide dismutase and glutathione peroxidase activities? // Biochem. Biophys. Res. Commun. – 2006. – V. 348. – P. 265-270.

24. De Kruijff B., Cullis P.R. Cytochrome c specifically induces non-bilayer structures in cardiolipin-containing model membranes // Biochim. Biophys. Acta. – 1980. – V. 602, I. 3. – P. 477-490.

25. Чупин В.В., Ушакова И.П., Бондаренко С.В., Василенко И.А., Серебренникова Г.А., Евстигнеева Р.П. Изучение взаимодействия метгемоглобина с модельными мембранами методом спектроскопии 31Р-ЯМР // Биоорг. Химия. – 1982. – Т. 8, N 9. – С. 1275-1280.
Cited
How to Cite
Kutsenko, O. K., Gorbenko, G. P., & Trusova, V. M. (1). Resonance energy transfer study of hemoglobin binding to model lipid membranes. Biophysical Bulletin, 1(24). Retrieved from https://periodicals.karazin.ua/biophysvisnyk/article/view/3895