Anesthetics Inhibit Acetylcholine-promoted Guanine Nucleotide Exchange of Heterotrimeric G Proteins of Airway Smooth Muscle

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Abstract

Background:

Anesthetics inhibit airway smooth muscle contraction in part by a direct effect on the smooth muscle cell. This study tested the hypothesis that the anesthetics halothane and hexanol, which both relax airway smooth muscle in vitro, inhibit acetylcholine-promoted nucleotide exchange at the α subunit of the Gq/11 heterotrimeric G protein (Gαq/11; i.e., they inhibit muscarinic receptor–Gαq/11 coupling).

Methods:

The effect of halothane (0.38 ± 0.02 mm) and hexanol (10 mm) on basal and acetylcholine-stimulated Gαq/11 guanosine nucleotide exchange was determined in membranes prepared from porcine tracheal smooth muscle. The nonhydrolyzable, radioactive form of guanosine-5′-triphosphate, [35S]GTPγS, was used as the reporter for Gαq/11 subunit dissociation from the membrane to soluble fraction, which was immunoprecipitated with rabbit polyclonal anti-Gαq/11 antiserum.

Results:

Acetylcholine caused a significant time- and concentration-dependent increase in the magnitude of Gαq/11 nucleotide exchange compared with basal values (i.e., without acetylcholine), reaching a maximal difference at 100 μm (35.9 ± 2.9 vs. 9.8 ± 1.2 fmol/mg protein, respectively). Whereas neither anesthetic had an effect on basal Gαq/11 nucleotide exchange, both halothane and hexanol significantly inhibited the increase in Gαq/11 nucleotide exchange produced by 30 μm acetylcholine (by 59% and 68%, respectively).

Conclusions:

Halothane and hexanol interact with the receptor–heterotrimeric G-protein complex in a manner that prevents acetylcholine-promoted exchange of guanosine-5′-triphosphate for guanosine-5′-diphosphate at Gαq/11. These data are consistent with the ability of anesthetics to interfere with cellular processes mediated by heterotrimeric G proteins in many cells, including effects on muscarinic receptor–G-protein regulation of airway smooth muscle contraction.

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