Junjie Wang, David George Jackson, Gerhard Dahl

The sequence and predicted membrane topology of Pannexin1 (Panx1) places it in the family of gap junction proteins. However, rather than forming gap junction channels, Panx1 forms channels in the non-junctional membrane. Panx1 operates in two distinct open states, depending on the mode of stimulation. The exclusively voltage-gated channel has a small conductance (<100 pS) and is highly selective for the flux of chloride ions. The Panx1 channel activated by various physiological stimuli or by increased concentrations of extracellular potassium ions has a large conductance (~500 pS, however with multiple, long lasting subconductance states) and is non-selectively permeable to small molecules, including ATP. To test whether the two open conformations also differ pharmacologically, the effects of di-and trivalent cations on the two Panx1 channel conformations were investigated. The rationale for this venture was that under certain experimental conditions ATP release from cells can be inhibited by multivalent cations, yet the literature indicates that the ATP release channel Panx1 is not affected by these ions. Consistent with previous reports, the Panx1 channel was not activated by removal of extracellular Ca2+ and the currents through the voltage-activated channel were not altered by Ca2+, Zn2+, Ba2+ or Gd3+. In contrast, the Panx1 channel activated to the large channel conformation by extracellular K+, osmotic stress or low oxygen was inhibited by the multivalent cations in a dose dependent way. Thus, monovalent cations activated the Panx1 channel from the closed state to the "large" conformation, while di- and tri-valent cations exclusively inhibited this large channel conformation.