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Effect of serum albumin and low molecular weight ligands on Zn uptake by isolated rat hepatocytes

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6962423
Although the liver plays an important role in Zn metabolism, the mechanisms regulating hepatic Zn uptake are poorly understood. Hepatocytes were isolated from adult female rats, body weight 200-250 g, by the collagenase perfusion technique and purified by differential centrifugation. Uptake of Zn was studied by incubation of hepatocytes with /sup 65/Zn in 10 mM HEPES buffer, pH 7.4, containing 138 mM NaCl, 5.2 mM KCl, 0.9 mM MgCl/sub 2/ and 1.0 mM CaCl/sub 2/, followed by washing with ice-cold 10 mM EDTA, 0.9% NaCl, pH 7.3 on Whatman GF-C glass microfiber filters. Zn uptake of 37/sup 0/C from ZnSO/sub 4/ (10 ..mu..M) was linear up to approx. 5 min with an initial rate of uptake of 11-20 ng/min/10/sup 6/ viable cells. Initial rate of Zn uptake at 37/sup 0/C was over 10-fold greater than at 4/sup 0/C. At 37/sup 0/C initial rate of uptake increased as a function of Zn concentration in the range 2-20 ..mu..M, with K/sub m/ = 27 ..mu..M and V/sub max/ = 42 ng/min/10/sup 6/ viable cells. Inclusion of bovine serum albumin (2 mg/ml) with ZnSO/sub 4/ (10 ..mu..m) reduced the initial rate of uptake to approx. 25% of the rate with ZnSO/sub 4/ alone. The low molecular weight ligands, phosphate, citrate and histidine at equimolar concentrations with Zn had little effect on Zn uptake by hepatocytes.
Research Organization:
Univ. College, Cork, Ireland
OSTI ID:
6962423
Report Number(s):
CONF-8604222-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:4
Country of Publication:
United States
Language:
English