Abstract
The mechanisms responsible for regulating epithelial ATP permeability and purinergic signaling are not well defined. Based on the observations that members of the ATP-binding cassette (ABC)1 family of proteins may contribute to ATP release, the purpose of these studies was to assess whether multidrug resistance-1 (MDR1) proteins are involved in ATP release from HTC hepatoma cells. Using a bioluminescence assay to detect extracellular ATP, increases in cell volume increased ATP release ̃3-fold. The MDR1 inhibitors cyclosporine A (10 μM) and verapramil (10 μM) inhibited ATP release by 69% and 62%, respectively (p < 0.001). Similarly, in whole-cell patch-clamp recordings, intracellular dialysis with C219 antibodies to inhibit MDR1 decreased ATP-dependent volume-sensitive Cl- current density from -33.1 ± 12.5 pA/pF to -2.0 ± 0.3 pA/pF (-80 reV, p 0.02). In contrast, overexpression of MDR1 in NIH 3T3 cells increased ATP release rates. Inhibition of ATP release by Gd3+ had no effect on transport of the MDR1 substrate rhodamine-123; and alteration of MDR1-substrate selectivity by mutation of G185 to V185 had no effect on ATP release. Since the effects of P-glycoproteins on ATP release can be dissociated from P-glycoprotein substrate transport, MDR1 is not likely to function as an ATP channel, but instead serves as a potent regulator of other cellular ATP transport pathways.
Original language | English (US) |
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Pages (from-to) | 165-173 |
Number of pages | 9 |
Journal | Journal of Membrane Biology |
Volume | 183 |
Issue number | 3 |
DOIs | |
State | Published - Oct 1 2001 |
Keywords
- ABC protein
- Cell volume
- Cl channel
- Liver
- Purinergic receptor
ASJC Scopus subject areas
- Biophysics
- Physiology
- Cell Biology