INVESTIGADORES
BARRANTES Francisco Jose
artículos
Título:
Changes in channel properties of acetylcholine receptors during the time course of thiol chemical modifications
Autor/es:
BOUZAT CB; BARRANTES FJ; SIGWORTH FJ
Revista:
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY
Editorial:
SPRINGER
Referencias:
Lugar: Berlin; Año: 1991 vol. 418 p. 51 - 61
ISSN:
0031-6768
Resumen:
The changes occurring during chemical modification of thiol groups in
single acetylcholine receptor (AChR) channels of BC3H-11 cells were
examined by the patch-clamp technique in the ?cell-attached?
configuration. Treatment with either 1 mM or 5 mM dithiothreitol or with
5 mM N-ethylmaleimide (NEM) does not cause
significant changes in the conductance and mean open time of the
channels. However, reduction with dithiothreitol followed by alkylation
with NEM produces modifications of AChRs. Under these conditions,
channels activated by 2 ìM acetylcholine show decreased open times
(about 15-fold shorter for the mostmodified AChRs) and a slight
reduction in single-channel current. Both changes are dependent on the
time of exposure and concentration of NEM. The rate of occurrence of
openings, however, changes little during NEM treatment. When reduced and
alkylated AChRs are activated by 100 ìM acetylcholine, clusters of
short openings separated by silent periods of about 1 s are observed.
The channel-open probability, determined for openings within a cluster,
is decreased by about 10-fold when compared with control receptors. The
observations at high agonist concentration indicate that the modified
AChR is still able to undergo desensitization.