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J. Biol. Chem., Vol. 262, Issue 28, 13483-13490, 10, 1987
F Okajima, Y Tokumitsu, Y Kondo and M Ui
Stimulation of P2-purinergic receptors by ATP resulted in activation of
phosphorylase, which was associated with marked production of inositol
trisphosphate (Ins-P3), in rat hepatocytes. ATP also inhibited
forskolin-induced accumulation of cAMP in the presence of a
phosphodiesterase inhibitor. On the contrary, adenosine or AMP never
inhibited the cAMP accumulation, but increased hepatocyte cAMP; the
stimulation was antagonized by a methylxanthine. Thus, P1-purinergic
receptors are linked to adenylate cyclase in a stimulatory fashion in
hepatocytes. Various kinds of purine nucleotides stimulating P2- receptors
can be divided into two groups on the basis of their relative abilities to
stimulate Ins-P3 production and to inhibit cAMP accumulation; the first
group including adenosine 5'-O-(3- thiotriphosphate) (ATP gamma S), ADP,
5-adenylyl imidodiphosphate, GTP, and guanosine 5'-O-(3-thiotriphosphate)
has an efficacy similar to that of ATP, and the second group of nucleotides
including alpha, beta- methyleneadenosine 5'-triphosphate, beta,
gamma-methyleneadenosine 5'- triphosphate (App(CH)2)p), and GDP exerts
considerable inhibitory effects on cAMP accumulation, but only slight
effects on inositol lipid metabolism. Treatment of hepatocytes with
islet-activating protein, pertussis toxin, blocked the nucleotide-induced
inhibition of cAMP accumulation, but exerted only a small effect on Ins-P3
production. In membranes prepared from hepatocytes, forskolin-stimulated
adenylate cyclase was inhibited by GTP. This GTP-induced inhibition of the
enzyme was susceptible to islet-activating protein and dependent on the
concentration of ATP (or its derivatives, ATP gamma S or App(CH2)p). It is
concluded that there are two types of P2-purinergic receptors: one is
linked to adenylate cyclase via an inhibitory guanine nucleotide regulatory
protein (Gi) and the other is linked to phospholipase C.
P2-purinergic receptors are coupled to two signal transduction systems leading to inhibition of cAMP generation and to production of inositol trisphosphate in rat hepatocytes
Department of Physical Biochemistry, Gunma University, Maebashi, Japan.
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