Expression of functional sodium channels from cloned cDNA.
The voltage-gated sodium channel is a transmembrane protein essential for the generation of action potentials in excitable cells. It has been reported that sodium channels purified from the electric organ of the electric eel, Electrophorus electricus, and from chick cardiac muscle consist of a single polypeptide of relative molecular mass (Mr) approximately 260,000, whereas those purified from rat brain and from rat and rabbit skeletal muscle contain, in addition to the large polypeptide, one or two smaller polypeptides of Mr 33,000-43,000. The primary structures of the Electrophorus sodium channel and two distinct sodium channel large polypeptides (designated as sodium channels I and II) from rat brain have been elucidated by cloning and sequencing the complementary DNAs. The purified sodium channel preparations from Electrophorus electroplax and from mammalian muscle and brain, when reconstituted into lipid vesicles or planar lipid bilayers, exhibit some functional activities. The successful reconstitution with the Electrophorus preparation would imply that the large polypeptide alone is sufficient to form functional sodium channels. However, studies with the rat brain preparation suggest that the smaller polypeptide of Mr 36,000 is also required for the integrity of the saxitoxin (STX) or tetrodotoxin (TTX) binding site of the sodium channel. Here we report that the messenger RNAs generated by transcription of the cloned cDNAs encoding the rat brain sodium channel large polypeptides, when injected into Xenopus oocytes, can direct the formation of functional sodium channels.
D000818:Animals; D004247:DNA / Q000235:genetics*; D004274:DNA, Recombinant / Q000502:physiology*; D005260:Female; D007473:Ion Channels / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D009865:Oocytes; D012333:RNA, Messenger / Q000302:isolation & purification / Q000502:physiology; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D015222:Sodium Channels; D014982:Xenopus laevis
D004274:DNA, Recombinant; D007473:Ion Channels; D008565:Membrane Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015222:Sodium Channels; D004247:DNA