STRUCTURAL-CHANGES IN PLASMA-MEMBRANES PREPARED FROM IRRADIATED CHINESE-HAMSTER V79-CELLS AS REVEALED BY RAMAN-SPECTROSCOPY

The effect of γ irradiation on the integrity of plasma membranes isolated from Chinese hamster V79 cells was investigated by Raman spectroscopy. Plasma membranes of control V79 cells show transitions between (-) 10 and 5°C (low-temperature transition), 10 and 22°C (middle-temperature transition), and 32 and 40°C (high-temperature transition). Irradiation (5 Gy) alters these transitions markedly. First, the low-temperature transition shifts to higher temperature (onset and completion temperatures 4 and 14°C). Second, the middle-temperature transition shifts up to the range of about 20-32°C, but the width remains unchanged. Third, the higher temperature transition broadens markedly and shifts to the range of about 15-40°C. Protein secondary structure as determined by least-squares analysis of the amide I bands shows 36% total helix, 55% total β-strand, and 9% turn plus undefined for control plasma membrane proteins. Plasma membrane proteins of irradiated V79 cells show an increase in total helix (40 and 45% at 5 and 10 Gy, respectively) and a decrease in the total β-strand (48 and 44% at 5 and 10 Gy, respectively) structures. The qualitative analysis of the Raman features of plasma membranes and model compounds in the 1600 cm-1 region, assigned to tyrosine groups, revealed that irradiation alters the microenvironment of these groups. We conclude that the radiation dose used in the survival range of Chinese hamster V79 cells can cause damage to plasma membrane proteins without detectable lipid peroxidation, and that the altered proteins react differently with lipids, yielding a shift in the thermal transition properties.