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The angiotensin II type 1 (AT1R) and type 2 (AT2R) receptors
belong to the seven transmembrane receptor superfamily. Previous
studies have suggested that the AT1R couples to a G
Volume 270,
Number 28,
Issue of July 14, pp. 16677-16682, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Coupling by the Use of Receptor Chimeras
signaling pathway, whereas the AT2R does not associate with
G. To identify the role that individual intracellular
domains play in AT1R function, AT1R/AT2R chimeric receptors were
prepared by substitution of intracellular loops. CHO cells expressing
these chimeras were used to test angiotensin II-induced c-fos expression and Ca mobilization which are
involved in the AT1R signaling pathway through G
coupling.
Substitution of the second intracellular loop (IC2) and the cytoplasmic
tail between the two receptors did not affect AT1R function. However,
exchange of the third intracellular loop (IC3) resulted in the loss of
function in the AT1R and conferred to the AT2R the ability to
constitutively activate the fos promoter. These findings
suggest that the third intracellular loop of the AT1R is critical for
G coupling. Substitution of discrete amino acid sequences
of the third intracellular loop indicate that its N-terminal and
C-terminal portions, especially the seven amino acids 219-225 in
the N-terminal portion, are important for AT1R function, and that the
intermediate portion of this loop is not required for G coupling.
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