Factors influencing the distribution pattern of porphyrins in cell membranes.
The mechanism of the sensitizer-membrane interactions has been studied by following the distribution properties of selected porphyrins, including haematoporphyrin (HP) and protoporphyrin (PP), into unilamellar liposomes of dipalmitoyl phosphatidylcholine (DPPC). The endomembrane distribution of HP and PP has been checked as a function of the membrane fluidity and composition by fluorescence polarization and quenching techniques. At porphyrin concentrations below 0.5 microM, HP and PP exclusively localize in the inner phospholipid monolayer; at higher concentrations, the outer monolayer also becomes populated. The porphyrin binding sites in liposomes, however, are different for HP and PP: HP preferentially distributes into water-accessible lipid regions, while PP localizes in the most hydrophobic loci of the lipid matrix. A porphyrin redistribution occurs when the fluidity properties of the liposomes are changed by addition of cholesterol or cardiolipin. In DPPC-cholesterol vesicles, all HP molecules dissolve in DPPC-rich regions while all PP molecules partition in cholesterol-rich environments. In DPPC-cardiolipin vesicles both porphyrins preferentially localize in regions accessible to the external medium. The effect of the nature of the carrier on porphyrin distribution in membranes has been studied by following the uptake and photosensitization properties of free and DPPC-incorporated PP and HP with rat liver mitochondria. The porphyrin photosensitizing efficiency has been checked by following the impairment of the respiratory function of mitochondria upon irradiation. Liposome-bound HP is less active than aqueous HP in determining membrane photodamage in mitochondria. On the contrary, aqueous PP is a very poor sensitizer as compared to a DPPC liposome-entrapped drug.(ABSTRACT TRUNCATED AT 250 WORDS)
Ricchelli
F
F
Department of Biology, University of Padova, Italy.
Jori
G
G
Moreno
G
G
Vinzens
F
F
Salet
C
C
D015060:1,2-Dipalmitoylphosphatidylcholine; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D008081:Liposomes; D008560:Membrane Fluidity; D011166:Porphyrins / Q000378:metabolism*; D011838:Radiation-Sensitizing Agents / Q000378:metabolism*
D008081:Liposomes; D011166:Porphyrins; D011838:Radiation-Sensitizing Agents; D015060:1,2-Dipalmitoylphosphatidylcholine