Ie. Kochevar et al., COMPARISON OF PHOTOSENSITIZED PLASMA-MEMBRANE DAMAGE CAUSED BY SINGLET OXYGEN AND FREE-RADICALS, Biochimica et biophysica acta. Biomembranes, 1280(2), 1996, pp. 223-230
The efficiency and selectivity of photosensitized damage to membrane f
unctions may be influenced strongly by the identity of the initial rea
ctive species formed by the photosensitizer. To test this possibility,
a phorosensitizer, rose bengal (RE), was used that resides in the pla
sma membrane and which generates singlet molecular oxygen (O-1(2).) up
on excitation with visible light, and radicals plus O-1(2). upon excit
ation with UV radiation. With this approach, O-1(2). and radicals are
formed at the same locations in the plasma membrane. The response of t
hree plasma membrane functions, namely, proline transport, membrane po
tential, and membrane impermeability to charged dye molecules, was ass
essed. The efficiencies of the responses in the presence and absence o
f oxygen were compared per photon absorbed by RE at two wavelengths; 3
55 nm (UV excitation) and 532 nm (visible excitation). The efficiency
of oxygen removal before irradiation was assessed by measuring the RE
tripler lifetime. The three membrane functions were inhibited more eff
iciently at 355 nm than at 532 nm in the presence of oxygen indicating
that the radicals are more effective at initiating damage to membrane
components than O-1(2). The ratio of photosensitized effects at the t
wo wavelengths in the presence of oxygen was the same for two membrane
functions but not for the third suggesting that O-1(2). and radicals
initiate a common mechanistic pathway for damage to some membrane func
tions but not to others. Removing oxygen reduced the efficiency of 355
nm-induced photosensitization by factors of 1.4 to 7. The sensitivity
of the three membrane functions to O-1(2).-initiated damage varied ov
er a factor of 50 whereas radical initiated damage only varied by a fa
ctor of 15. In summary, these results indicate that radicals and O-1(2
). formed at the same locations in the plasma membrane vary in their e
fficiency and specificity for membrane damage but may, in some cases,
operate by a common secondary damage mechanism in the presence of oxyg
en.