Transport by the (Na<sup>+</sup>, K<sup>+</sup>) ATPase: Modulation by differentiation inducers and inhibition of protein synthesis in the MDCK kidney epithelial cell line

Journal of Cellular Physiology - Tập 123 Số 3 - Trang 410-416 - 1985
Brian G. Kennedy1, Julia E. Lever1
1Department of Biochemistry and Molecular Biology, The University of Texas Medical School, Houston, Texas 77225

Tóm tắt

AbstractMDCK kidney epithelial cell cultures exposed to the differentiation inducer hexamethylene bisacetamid (HMBA) for 24 hours exhibited a 50% decrease in transport activity per (Na+, K+)‐ATPase molecule (turnover number) but an unchanged number of pump sites (Kennedy and Lever, 1984). Inhibition of protein synthesis by either 10 μM cycloheximide or 2 μM emetine blocked the inhibitory effects of HMBA on Na+/K+ pump efficiency assessed by measurements of [3H]‐ouabain binding to intact cells, (Na+, K+) ATPase activity of detergent‐activated cell extracts, and ouabain‐sensitive Rb+ uptake. In the absence of inducer treatment, inhibition of protein synthesis increased Na+/K+ pump turnover number by twofold while maintaining Na+/K+ pump activity per cell at a constant level. Intracellular Na+ levels were decreased after cycloheximide treatment; therefore, pump stimulation was not due to substrate effects. Furthermore, cycloheximide effects of Rb+ uptake could be dissociated from effects on tight junctions. These observations suggest that the transport activity of the (Na+, K+) ATPase is tightly regulated by factors dependent on protein synthesis.

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