Abstract
Fluorescence spectra and fluorescence lifetimes of protochlorophyll (Pchl) were measured in organic solvents having different physical and chemical properties and were analyzed taking into account the nonspecific (dependent on bulk solvent parameters), and specific (e.g. H bonds, Mg coordination) solvent–solute interactions. The energy of the fluorescence emission band decreased, while the Stokes shift increased for increasing solvent orientation polarizability, which is a function of both the dielectric constant (ε) and the refractive index (n). The extent of the dependence of the Stokes shift on solvent orientation polarizability was higher in protic (i.e. those able to form hydrogen-binding) than in aprotic solvents. High value of the Stokes shift was also observed in pyridine and methanol, i.e. in solvents hexacoordinating the central Mg atom. The fluorescence decay of Pchl was monoexponential in all of the investigated solvents. The fluorescence lifetime decreased for increasing solvent orientation polarizability from 5.5 ± 0.1 ns in 1,4-dioxane to 3.3 ± 0.1 ns in methanol. Longer lifetime values were observed in the case of aprotic solvents than in protic solvents. The hexacoordination of Mg had no effect on the fluorescence lifetime. The present data are discussed with respect to results found for protochlorophyllide (Pchlide) (Myśliwa-Kurdziel et al. in Photochem Photobiol 79:62–67, 2004), and they indicate that the presence of phytol chain in the porphyrin ring influences the spectral properties of the whole chromophore. This is the first complex analysis comparing the fluorescence emission and fluorescence lifetimes of purified Pchl and Pchlide.
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Abbreviations
- Bchl:
-
Bacteriochlorophyll
- Bchlide:
-
Bacteriochlorophyllide
- Chl:
-
Chlorophyll
- Chlide:
-
Chlorophyllide
- cP:
-
Centipoises
- DMSO:
-
Dimethyl sulfoxide
- DV-:
-
Divinyl
- EPIM:
-
Etioplasts inner membranes
- GG:
-
Geranylgeraniol
- MV-:
-
Monovinyl
- Pchlide:
-
Protochlorophyllide
- Pchl:
-
Protochlorophyll
- Pchl(ide):
-
Mixture of Pchl and Pchlide of unknown proportions
- THF:
-
Tetrahydrofuran
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Acknowledgments
This work was financed from the budget of the Polish Ministry of Science for the years 2005–2008 under the project No: 2 P04A 063 28. B. M.-K. was the recipient of a Scholarship of the Rector of the Jagiellonian University in the academic years 2004–2006. Anna Cwanek MSc is kindly acknowledged for her technical help in part of the fluorescence lifetime measurements.
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Regional Biophysics Conference of the National Biophysical Societies of Austria, Croatia, Hungary, Italy, Serbia, and Slovenia.
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Myśliwa-Kurdziel, B., Solymosi, K., Kruk, J. et al. Solvent effects on fluorescence properties of protochlorophyll and its derivatives with various porphyrin side chains. Eur Biophys J 37, 1185–1193 (2008). https://doi.org/10.1007/s00249-008-0288-x
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DOI: https://doi.org/10.1007/s00249-008-0288-x