Modification of transmembrane electron transport activity in plasma membranes of simian virus 40 transformed pineal cells.
Changes have been found in the plasma membrane enzyme system which carries out transmembrane electron transport and associated proton transport in Simian virus 40 (SV40) temperature-sensitive A (tsA) mutant-transformed rat pineal cell line, RPN209-1. This cell line was temperature-sensitive for the maintenance of transformation. RPN209-1 cells expressed the transformed phenotype (rapid growth, high cell density, and cloning in soft agar) at the permissive temperature (33 degrees C) and the nontransformed phenotype (slower growth, lower saturation density, and lower cloning efficiency in soft agar) at the nonpermissive temperature (40 degrees C). The reduction of external ferricyanide, hexaammine ruthenium and diferric transferrin was used to measure the transmembrane redox activity. The transformed RPN209-1 cells expressed a lower transmembrane redox activity, which is more sensitive to the antitumor drug adriamycin, when compared to the cells with a nontransformed phenotype. The lower transmembrane redox activity is associated with a decrease in the affinity for ferricyanide and a change in Vmax of the enzyme. Since the transformed cells have 25% lower concentration of NADH, the decrease in Vmax may be partly based on substrate limitation. Ionic strength variation in the assay media shows that the change in activity with transformation is not based on change in cell-surface change. Treatment with neuraminidase, however, indicates that sialic acid is important for enzyme activity, consistent with previous proposals that the transmembrane enzyme is a glycoprotein. The proton extrusion associated with transplasma membrane electron transport is increased in transformed cells relative to the rate of ferricyanide reduction. A relation between proton pumping transplasma membrane electron transport and growth stimulation by external oxidants is discussed.
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002472:Cell Transformation, Viral; D004579:Electron Transport / Q000187:drug effects; D005292:Ferricyanides / Q000378:metabolism; D009439:Neuraminidase / Q000494:pharmacology; D010084:Oxidation-Reduction; D010870:Pineal Gland / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D013539:Simian virus 40 / Q000378:metabolism*; D013696:Temperature
D005292:Ferricyanides; C007931:hexacyanoferrate III; D009439:Neuraminidase