TITYUSTOXIN K-ALPHA BLOCKS VOLTAGE-GATED NONINACTIVATING K-DENDROTOXIN IN SYNAPTOSOMES( CHANNELS AND UNBLOCKS INACTIVATING K+ CHANNELS BLOCKED BY ALPHA)

Two nonhomologous polypeptide toxins, tityustoxin K alpha(TsTX-K alpha ) and tityustoxin K beta(TsTX-K beta), purified from the venom of the Brazilian scorpion Tityus serrulatus, selectively block voltage-gated noninactivating K+ channels in synaptosomes (IC50 values of 8 nM and 3 0 nM, respectively). In contrast, alpha-dendrotoxin (alpha-DTX) and ch arybdotoxin (ChTX) block voltage-gated inactivating K+ channels in syn aptosomes (IC50 values of 90 nM and 40 nM, respectively). We studied i nteractions among these toxins in I-125-alpha-DTX binding and Rb-86 ef flux experiments. Both TsTX-K alpha and ChTX completely displaced spec ifically bound I-125-alpha-DTX from synaptic membranes, but TsTX-K bet a had no effect on bound alpha-DTX. TsTX-K alpha and TsTX-K beta block ed the same noninactivating component of 100 mM K+-stimulated Rb-86 ef flux in synaptosomes. Both alpha-DTX and ChTX blocked the same inactiv ating component of the K+-stimulated Rb-86 efflux in synaptosomes. Bot h the inactivating and the noninactivating components of tile 100 mM K +-stimulated Rb-86 efflux were completely blocked when 200 nM TsTX-K b eta and either 600 nM alpha-DTX or 200 nM ChTX were present. The effec ts of TsTX-K alpha and ChTX on Rb-86 efflux were also additive. When T sTX-K alpha was added in the presence of alpha-DTX, however, only the noninactivating component of the K+-stimulated efflux was blocked. The inactivating component could then be blocked by ChTX, which is struct urally homologous to TsTX-K alpha. We conclude that TsTX-K alpha unblo cks the voltage-gated inactivating K+ channels in synaptosomes when th ey are blocked by alpha-DTX, but not when they are blocked by ChTX. Ts TX-K alpha binds to a site on the inactivating K+ channel that does no t occlude the pore; its binding apparently prevents alpha-DTX (7054 Da ), but not ChTX (4300 Da), from blocking the pore. The effects of TsTX -K alpha on I-125-alpha-DTX binding and Rb-86 efflux are mimicked by n oxiustoxin, which is homologous to TsTX-K alpha and ChTX.