Hm. Clawges et al., HUMAN 5-HT1 RECEPTOR SUBTYPES EXHIBIT DISTINCT G-PROTEIN COUPLING BEHAVIORS IN MEMBRANES FROM SF9 CELLS, Biochemistry, 36(42), 1997, pp. 12930-12938
The G protein coupling behavior of four human 5-hydroxytryptamine rece
ptor subtypes (5-HT1A, 5-HT1B, 5-HT1D, and 5-HT1E) has been studied in
membranes from Sf9 cells expressing the individual receptors. The 5-H
T1A and 5-HT1B receptors exhibited both high-and low-affinity states f
or agonist, with the majority of the receptors in a low-affinity state
. Addition of purified G protein subunits to membranes expressing eith
er 5-HT1A or 5-HT1B receptors shifted the majority of the receptors to
a high-affinity state in the absence, but not in the presence, of gua
nine nucleotides. The alpha(i1), alpha(i2), alpha(i3), and alpha(0) su
bunits were able to shift the receptors to a high-affinity state with
either beta gamma(brain) or beta gamma(retina) while alpha(t) subunits
were inactive regardless of which beta gamma preparation was used. A
significantly higher affinity for agonist was observed with both recep
tors in the presence of alpha(i3) subunits compared with either alpha(
i2) or alpha(0) subunits, while a significantly lower concentration of
alpha subunits was required for a maximal affinity shift of 5-HT1A re
ceptors compared with 5-HT1B receptors (EC50 values of 6.4 and 12.0 nM
, respectively). The 5-HT1D and 5-HT1E receptors exhibited only a sing
le affinity state for agonist. Addition of purified G protein subunits
to membranes containing 5-HT1D receptors caused a small increase in a
ffinity for agonist that was only partially reversed by guanine nucleo
tides while the addition of purified G protein subunits to membranes c
ontaining 5-HT1E receptors had no affect on agonist binding. Thus when
expressed in an identical membrane environment these four closely rel
ated 5-HT1 receptor subtypes exhibit different G protein coupling beha
viors.