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Virol J
2008 May 19;5:60. doi: 10.1186/1743-422X-5-60.
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Alterations in intracellular potassium concentration by HIV-1 and SIV Nef.
Choi B
,
Fermin CD
,
Comardelle AM
,
Haislip AM
,
Voss TG
,
Garry RF
.
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HIV-1 mediated perturbation of the plasma membrane can produce an alteration in the transmembrane gradients of cations and other small molecules leading to cell death. Several HIV-1 proteins have been shown to perturb membrane permeability and ion transport. Xenopus laevis oocytes have few functional endogenous ion channels, and have proven useful as a system to examine direct effects of exogenously added proteins on ion transport. HIV-1 Nef induces alterations in the intracellular potassium concentration in CD4+ T-lymphoblastoid cells, but not intracellular pH. Two electrode voltage-clamp recording was used to determine that Nef did not form ion channel-like pores in Xenopus oocytes. These results suggest that HIV-1 Nef regulates intracellular ion concentrations indirectly, and may interact with membrane proteins such as ion channels to modify their electrical properties.
Figure 1. Alterations in intracellular K+ in T-lymphoblastoid cells incubated with recombinant HIV-1 and SIV Nef protein. H9 cells were loaded either with the K+ sensitive fluorescent indicated PBFI-AM (panel A) or the pH sensitive fluorescent indicator BCECF-AM (Panel B), then incubated with various concentrations of recombinant HIV-1 or SIV Nef protein for 15 min at 37°C, and fluorescence intensity was measured by using a fluorescence concentration analyzer. Each data point represents the mean ± standard error of eight independent determinations.
Figure 2. Fluorescent and light microscopic analysis of alteration in intracellular K+ concentrations in H9 cells treated with recombinant Nef. H9 cells were loaded with the fluorescent indicator PBFI-AM and incubated with 300 nM recombinant Nef protein or with control medium without Nef for 15 min at 37°C. Control cells (A and C) and cells incubated with Nef protein (B and D) were examined by light (A and B) and fluorescent (C and D) microscopy.
Figure 3. Membrane currents in Xenopus oocytes exposed to Nef protein. (A) 300 nM recombinant Nef, (B) control oocyte Panel C: current:voltage plots determined from data recorded in panel A and B.
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