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AJP - Endocrinology and Metabolism, Vol 250, Issue 6 677-E685, Copyright © 1986 by American Physiological Society
ARTICLES |
S. E. Pattison and R. J. Cousins
The kinetics of 65Zn2+ uptake and exchange by hepatocytes in primary culture have been examined in detail to provide a basis for analyzing hormonal regulation of hepatic zinc metabolism. 65Zn2+ uptake was found to be a biphasic process. The slow phase represents an exchange between Zn2+ in the medium and preexisting, intracellular zinc pools. This exchange rate was saturable with a medium zinc concentration of 9.5 microM eliciting one-half the maximum exchange rate and a maximum exchange rate of 9.9 pmol Zn2+ . min-1 . mg protein-1 in the presence of bovine serum albumin. In the absence of albumin, a secondary, nonsaturable uptake rate was observed. The slow phase was relatively selective, and of the divalent transition metal ions tested, only Cd2+ and Mn2+ caused inhibition. The rate of exchange suggests total hepatocyte zinc has a turnover rate of approximately 30 h. The fast phase of 65Zn2+ reflects net Zn2+ accumulation into a labile pool. The initial rates for this process were too fast to be measured accurately, but steady-state measurements allowed determination of the labile pool size. The pool dimensions saturated in the presence [Kapp = 28.6 microM; pool capacity = 0.44 nmol Zn2+/mg protein] and absence [Kapp = 11.8 microM; pool capacity = 0.34 nmol Zn2+/mg protein] of bovine serum albumin. Kinetics and equilibria of Zn2+ uptake into the labile pool suggest that the latter acts as a source of Zn2+ for the slow-exchange phase. Dexamethasone stimulated slow Zn2+ exchange and also increased the labile pool size. The data suggest physiological factors alter hepatic zinc metabolism by influencing both intracellular Zn2+ pools.
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