RT Journal Article
SR Electronic
T1 Baculovirus Expression Provides Direct Evidence for Heteromeric Assembly of P2X2 and P2X3 Receptors
JF The Journal of Neuroscience
JO J. Neurosci.
FD Society for Neuroscience
SP 6529
OP 6533
DO 10.1523/JNEUROSCI.17-17-06529.1997
VO 17
IS 17
A1 Radford, Kathryn M.
A1 Virginio, Caterina
A1 Surprenant, Annmarie
A1 North, R. Alan
A1 Kawashima, Eric
YR 1997
UL http://www.jneurosci.org/content/17/17/6529.abstract
AB P2X2 and P2X3 are subunits of P2X receptors, cation channels opened by binding extracellular ATP. cDNAs encoding P2X2 and P2X3 receptor subunits, each with one of two C-terminal epitope tags, were cloned into baculovirus. Virally infected insect cells (Spodoptera frugiperda) expressed moderate to high levels of the corresponding proteins, as detected by Western blotting, by the specific binding of [35S]ATP and by whole-cell recordings of membrane current evoked by ATP or αβmethylene-ATP. In cells infected at the same time with two viruses encoding P2X2 and P2X3 receptors, the two proteins could be cross-immunoprecipitated with antibodies specific for either of the epitope tags. Whole-cell recordings from these cells showed that ATP and αβmethylene-ATP evoked currents with agonist sensitivity and desensitization quite distinct from those observed when P2X2 or P2X3 receptors were expressed alone. The results offer a method to express large amounts of P2X receptor protein, and they provide direct evidence that P2X2 and P2X3 subunits assemble to form heteromeric channels having distinct properties from those formed as homomers.