Single-channel currents of the permeability transition pore from the inner mitochondrial membrane of rat liver and of a human hepatoma cell line.
Single-channel currents were recorded from inner mitochondrial membranes of HepG2 hepatoma cells and of normal rat liver cells by means of patch-clamp techniques. The current events showed variable amplitudes of up to 1.1 +/- 0.2 nS (n = 35) at room temperature (24 degrees C) and of up to 1.5 +/- 0.2 nS (n = 10) at 34 degrees C including large numbers of subconductance states. Voltages of -40 mV and below closed the channels usually with a delay of about 2 min. Increasing Ca(2+) concentrations activated the channels, whereas cyclosporin A (100 nM) blocked. The concentration-response relation for the Ca(2+)-effect could be fitted best using an EC(50) of 10 microM and a Hill coefficient of 1.5. Taken together the results indicate that we recorded from the permeability transition pore (PTP). As PTP activity may be related to apoptosis we tested if lysate from differently treated T-lymphocytes (Jurkat cells) was able to induce PTP activity in HepG2 cells. Lysate of untreated cells completely abolished the activity at a Ca(2+) concentration of 18 nM (buffered by EGTA), i.e. three orders of magnitude below the EC(50). Under these conditions the lysate is not likely to contain stable factors that could open the PTP. Preliminary experiments show PTP activity in CD95-activated lysate.
Loupatatzis
Christos
C
Institut für Biochemie, Universität Magdeburg, Germany.
Seitz
Gordon
G
Schönfeld
Peter
P
Lang
Florian
F
Siemen
Detlef
D
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002118:Calcium / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D016572:Cyclosporine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D006801:Humans; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D019169:Jurkat Cells; D008113:Liver Neoplasms / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D018408:Patch-Clamp Techniques; D051381:Rats; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000494:pharmacology
D007473:Ion Channels; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D019014:fas Receptor; D016572:Cyclosporine; D002118:Calcium