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Vol. 284, Issue 3, 1218-1226, March 1998
hm5
Subtypes: Effect of G Protein-Coupled Receptor Kinases GRK2, GRK4, GRK5
and GRK61
Department of Neurochemistry, Faculty of Medicine, The University
of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113, Japan
Sequestration of porcine muscarinic acetylcholine receptor m2 subtypes
(m2 receptors) expressed in COS-7 cells is facilitated by coexpression
of G protein-coupled receptor kinases 2 (GRK2). We examined the effect
of coexpression of GRK2, GRK4
, GRK5 and GRK6 on sequestration of
human m1
m5 receptors expressed in COS-7 cells, which was assessed as
loss of [3H]N-methylscopolamine binding activity from the
cell surface. Sequestration of m4 receptors as well as m2 receptors was
facilitated by coexpression of GRK2 and attenuated by coexpression of
the dominant negative form of GRK2 (DN-GRK2). Sequestration of m3 and
m5 receptors also was facilitated by coexpression of GRK2 but not
affected by coexpression of DN-GRK2. On the other hand, proportions of
sequestered m1 receptors were not significantly different with
coexpression of GRK2 and DN-GRK2. GRK4, GRK5 and GRK6 did not
facilitate sequestration of m1m5 receptors in COS-7 cells, except
that the sequestration of m2 receptors tended to be facilitated by
coexpression of GRK4, GRK5 and GRK6. However, coexpression of
GRK4, GRK5, but not GRK6, in BHK-21 cells facilitated sequestration
of m2, but not m3, receptors. These results indicate that the effect of
GRK2 to facilitate receptor sequestration is not restricted to m2
receptors but is generalized to other muscarinic receptors except m1
receptors and that other kinases, including GRK4, GRK5 and
endogenous kinase(s) in COS-7 cells, also contribute to sequestration
of m2 and m4 receptors.
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