Inactivation of cloned Na channels expressed in Xenopus oocytes.
This study investigates the inactivation properties of Na channels expressed in Xenopus oocytes from two rat IIA Na channel cDNA clones differing by a single amino acid residue. Although the two cDNAs encode Na channels with substantially different activation properties (Auld, V. J., A. L. Goldin, D. S. Krafte, J. Marshall, J. M. Dunn, W. A. Catterall, H. A. Lester, N. Davidson, and R. J. Dunn. 1988. Neuron. 1:449-461), their inactivation properties resemble each other strongly but differ markedly from channels induced by poly(A+) rat brain RNA. Rat IIA currents inactivate more slowly, recover from inactivation more slowly, and display a steady-state voltage dependence that is shifted to more positive potentials. The macroscopic inactivation process for poly(A+) Na channels is defined by a single exponential time course; that for rat IIA channels displays two exponential components. At the single-channel level these differences in inactivation occur because rat IIA channels reopen several times during a depolarizing pulse; poly(A+) channels do not. Repetitive stimulation (greater than 1 Hz) produces a marked decrement in the rat IIA peak current and changes the waveform of the currents. When low molecular weight RNA is coinjected with rat IIA RNA, these inactivation properties are restored to those that characterize poly(A+) channels. Slow inactivation is similar for rat IIA and poly(A+) channels, however. The data suggest that activation and inactivation involve at least partially distinct regions of the channel protein.
Division of Biology, California Institute of Technology, Pasadena 91125.
D S
DS
Krafte
A L
AL
Goldin
V J
VJ
Auld
R J
RJ
Dunn
N
N
Davidson
H A
HA
Lester
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005260:Female; D066298:In Vitro Techniques; D007700:Kinetics; D008564:Membrane Potentials; D009865:Oocytes / Q000378:metabolism; D011061:Poly A / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger; D051381:Rats; D015222:Sodium Channels / Q000235:genetics / Q000378:metabolism*; D014981:Xenopus
D012333:RNA, Messenger; D015222:Sodium Channels; D011061:Poly A; D012313:RNA; D004247:DNA