A fluorescence study of the interaction and location of (+)-totarol, a diterpenoid bioactive molecule, in model membranes.
(+)-Totarol, a diterpene extracted from Podocarpus totara, has been reported as a potent antioxidant and antibacterial agent. Although the molecular mechanism of action of this hydrophobic molecule remains unknown, recent work made in our laboratory strongly suggests that it could be lipid-mediated. Since (+)-totarol contains a phenolic ring, we have studied the intrinsic fluorescent properties of this molecule, i.e., quantum yield, lifetime, steady-state anisotropy and emission spectra, both in aqueous and in phospholipid phases, in order to obtain information on the interaction and location of (+)-totarol in biomembrane model systems. The phospholipid/water partition coefficient of (+)-totarol was found to be very high (K(p)=1.8x10(4)), suggesting that it incorporates very efficiently into membranes. In order to estimate the transverse location (degree of penetration) of the molecule in the fluid phase of DMPC model membranes, the spin labelled fatty acids 5-NS and 16-NS were used in differential quenching experiments. The results obtained show that (+)-totarol is located in the inner region of the membrane, far away from the phospholipid/water interface. Since (+)-totarol protects against oxidative stress, its interaction with an unsaturated fatty acid, trans-parinaric acid, was studied using fluorescence resonance energy transfer. No significant interactions were observed, molecules of trans-parinaric acid distributing themselves randomly amongst those of (+)-totarol in the phospholipid membrane.
Mateo
C R
CR
Centro de Biología Molecular y Celular, Universidad Miguel Hernández, Elche-Alicante, Spain.
Prieto
M
M
Micol
V
V
Shapiro
S
S
Villalaín
J
J
D045784:Abietanes; D016880:Anisotropy; D004134:Dimyristoylphosphatidylcholine; D004224:Diterpenes / Q000737:chemistry*; D004735:Energy Transfer; D005231:Fatty Acids, Unsaturated / Q000737:chemistry; D005456:Fluorescent Dyes; D008051:Lipid Bilayers / Q000737:chemistry*; D008566:Membranes / Q000737:chemistry*; D015394:Molecular Structure; D010743:Phospholipids / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013696:Temperature
D045784:Abietanes; D004224:Diterpenes; D005231:Fatty Acids, Unsaturated; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D010743:Phospholipids; C078548:totarol; C013641:parinaric acid; D004134:Dimyristoylphosphatidylcholine