Rb+ influx in response to changes in energy generation: effect of the regulation of the ATP content of HeLa cells.

Abstract

The effect of changes in energy metabolism on Rb+ influx was studied in HeLa cells. Irrespective of whether ATP production was controlled by varying the activity of glycolysis or of oxidative metabolism on addition of certain combinations of glucose, carbonylcyanide m-chlorophenylhydrazone, monoiodoacetic acid, and quercetin, Rb+ influx changed as a linear function of the ATP content, which varied in a wide range up to the normal level (15-20 nmol/mg protein or 3-4 mM). The difference between results obtained by these procedures was not significant. As the intracellular Na+ content varied at different ATP contents, the Na+ content was adjusted to similar levels by chilling the cells with varying ATP contents. However, a linear relation was still observed. A similar dependence was also obtained for cytoplasmic ATP, which would be more closely connected with the Na,K-pump than total ATP. The ratio of ouabain-sensitive Rb+ influx to the corresponding part of lactate production was nearly 2 in the presence of 2 mM glucose. From these results it is concluded that (1) active Rb+ influx, which is chiefly maintained by energy generated through glycolysis, can also be supported by oxidative metabolism; (2) Rb+ influx is regulated linearly as a function of the cellular ATP content up to the control level; but does not increase if ATP is raised still further; and (3) 2 Rb+ ions move concomitantly at the expense of one ATP molecule.