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. Author manuscript; available in PMC: 2009 Jan 23.
Published in final edited form as: FEBS Lett. 2007 Dec 26;582(2):299–304. doi: 10.1016/j.febslet.2007.12.022

Table 1.

Enhancement of selenium uptake by thiols in HaCat cells demonstrates evidence for high affinity uptake of selenide

Rxn Tested compoundsa Selenium retained
(picomoles)		 Selenium retainedb
of total (%)
1 selenite (control) 0.066 ± 0.011 0.36 ± 0.063
2 selenite, 10μM glutathione 0.50 ± 0.12 2.8 ± 0.66
3 selenite, 10μM dithiothreitol 5.3 ± 0.64 29 ±3.6
4 selenite, 10μM β-mercatoethanol 4.0 ± 0.46 22 ± 2.6
5 selenite, 10μM Trx 0.071 ±0.02 0.39 ± 0.11
6 selenite, 15μM NADPH 0.062 ± 0.015 0.34 ± 0.085
7 selenite, 10μM Trx, 15μM NADPH 0.076 ± 0.018 0.42 ± 0.099
8 selenite, TrxR, Trx, NAPDH 1.8 ± 0.28 10 ± 1.6
a

Thiols were pre-incubated with selenite for 10 minutes and then entire mixture was added to HaCat cells containing 1 mL of DPBS and incubated for 1 hour. Reactions 5-8 contained either 1 μM of thioredoxin, 1.5 μg of thioredoxin reductase or 500 μM of NADPH in a final volume of 30 μL, with only reaction 8 containing all the components to regenerate reduced thioredoxin catalytically.

b

Total selenium retained was determined after several washes of the HaCat cell monolayer and subsequent lysis by NaOH as described in methods. The mean of two independent experiments (with multiple replicates) is shown with standard deviation as the error.