Effect of Me(2)SO on membrane phase behavior and protein denaturation of human pulmonary endothelial cells studied by in situ FTIR spectroscopy.
Fourier transform infrared spectroscopy (FTIR) provides a unique technique to study membranes and proteins within their native cellular environment. FTIR was used here to study the effects of dimethyl sulfoxide (Me(2)SO) on membranes and proteins in human pulmonary endothelial cells (HPMECs). Temperature-dependent changes in characteristic lipid and protein vibrational bands were identified to reveal the effects of Me(2)SO on membrane phase behavior and protein stability. At Me(2)SO concentrations equal to or below 10% (v/v), Me(2)SO was found to decrease membrane conformational disorder. At higher Me(2)SO concentrations (15% v/v), however, membrane conformational disorder was found to be similar to that of cells in the absence of Me(2)SO. This effect was observed over a wide temperature range from 90 degrees C down to -40 degrees C. Me(2)SO had no clear effects on cellular proteins during freezing. During heating, however, Me(2)SO had a destabilizing effect on cellular proteins. In the absence of Me(2)SO, protein denaturation started at an onset temperature of 46 degrees C, whereas at 15% Me(2)SO the onset temperature of protein denaturation decreased to 32 degrees C. This implies that in the presence of Me(2)SO the onset temperature of protein denaturation is lower than the normal growth temperature of the cells, which could explain the well documented toxic effect of Me(2)SO at physiological temperatures. Me(2)SO destabilizes cellular proteins during heating and decreases membrane conformational disorder over a wide temperature range.
Spindler
Ralf
R
Institute of Multiphase Processes and Centre for Biomedical Engineering, Leibniz Universitaet Hannover, Hannover, Germany. spindler@ifv.uni-hannover.de
Wolkers
Willem F
WF
Glasmacher
Birgit
B
D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D044367:Phase Transition / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D011506:Proteins / Q000378:metabolism*; D011651:Pulmonary Artery / Q000187:drug effects / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D003451:Cryoprotective Agents; D011506:Proteins; D004121:Dimethyl Sulfoxide