The role of protein kinase A and protein kinase C in prostanoid IP receptor desensitization in NG108-15 cells

Krane, Anke; Malkhandi, Joy; Mercy, Linda; Keen, Mary

doi:10.1016/0167-4838(94)90209-7

Cited by 9 publications

(7 citation statements)

References 16 publications

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“…FURA2/AM was purchased from Calbiochem. [ 3 H] i P 3 (20 -40 Ci/mmol) and [ 3 H]cAMP (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30) Ci/mmol) were purchased from American Radiolabeled Chemicals Inc. [ 3 H]Iloprost (15.3Ci/mmol) was purchased from Amersham Biosciences. [9,10-3 H]Palmitic acid (60 Ci/mmol) was obtained from PerkinElmer Life Sciences.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, it was established that in addition to G s , the mouse IP also couples to inhibition of adenylyl cyclase via G i and to G q /PLC activation through a switching mechanism involving its initial coupling to G s /adenylyl cyclase activation and subsequent cAMP-dependent protein kinase A phosphorylation of the mouse IP at Ser 357 within its C-tail region (20). Additionally, IPs are widely reported to undergo rapid agonist-induced desensitization, internalization, and down-regulation in human platelets and in other cell types (21)(22)(23)(24).…”

Section: Superfamily (1 2 4 5)mentioning

confidence: 99%

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Palmitoylation of the Human Prostacyclin Receptor

Miggin

Lawler

Kinsella

2003

Journal of Biological Chemistry

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, nor hIP C308S,C309S,C311S coupled to G q . Taken together, these data confirm that the hIP is isoprenylated and palmitoylated, and collectively these modifications modulate its G protein coupling and effector signaling. We propose that through lipid modification followed by membrane insertion, the C-tail domain of the IP may contain a double loop structure anchored by the dynamically regulated palmitoyl groups proximal to transmembrane domain 7 and by a distal farnesyl isoprenoid permanently attached to its carboxyl terminus.Modification of proteins through the covalent attachment of lipid groups occurs within a wide variety of cellular proteins and may be involved in mediating protein-membrane and/or protein-protein interactions (1-3). Three of the most common lipid modifications are N-myristoylation, isoprenylation and thio(S)-acylation. In contrast to myristoylation and isoprenylation, which typically occur either as co-translational or as immediate post-translational events, S-acylation through attachment of palmitate to Cys residue(s), via a labile thioester bond, occurs post-translationally (1). Moreover, whereas the former two modifications remain attached until protein degradation, palmitoylation is a reversible, dynamic modification that turns over more rapidly than the protein itself and thus has the potential to be regulated (2, 3). A diverse family of cellular proteins are established to be palmitoylated including ␣ subunits of the heterotrimeric G protein subunits, for example G␣ s and G␣ q , Ha-and N-Ras proteins, A kinase anchoring protein 15 and 18, endothelial nitric-oxide synthase, adenylyl cyclase, G protein-coupled receptor kinases 4 and 6, diverse members of the Src family of nonreceptor tyrosine kinases, in addition to several members of the G protein-coupled receptor (GPCR)

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Section: Methodsmentioning

confidence: 99%

Section: Superfamily (1 2 4 5)mentioning

confidence: 99%

Palmitoylation of the Human Prostacyclin Receptor

Miggin

Lawler

Kinsella

2003

Journal of Biological Chemistry

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“…Instead, short-term desensitization of these receptors is thought to occur by homologous mechanisms involving GRK2 . Prostanoid-IP receptors are proposed to desensitize via down-regulation of the receptor protein and G sa from the cell surface in NG108-15 cells, without the involvement of PKC (Krane et al, 1994;Williams & Kelly, 1994). In the present study, the lack of e ect of PKC activation on desensitization of A 2 adenosine and prostanoid-IP receptors was therefore consistent with previous reports.…”

Section: British Journal Of Pharmacology Vol 135 (8)supporting

confidence: 93%

“…Also, the present study suggests that PKA is selectively regulating secretin receptor signalling at the level of the receptor itself, as adenylyl cyclase responses to forskolin, iloprost and NECA were una ected by PKA inhibition. It has been reported that forskolin-mediated PKA activation can heterologously regulate A 2 adenosine and prostanoid IP receptor desensitization, however, these e ects were only observed after 17 h of forskolin treatment and were consistent with a decrease in receptor expression (Keen et al, 1992;Krane et al, 1994). As secretin partly exerts its physiological e ects via stimulation of adenylyl cyclase with subsequent activation of PKA (Ulrich et al, 1998), feedback inhibition by PKA is likely to play an important role in physiological regulation of secretin receptor responsiveness.…”

Section: British Journal Of Pharmacology Vol 135 (8)mentioning

confidence: 93%

Second messenger‐dependent protein kinases and protein synthesis regulate endogenous secretin receptor responsiveness

Ghadessy

Kelly

2002

British J Pharmacology

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1 The present study investigated the role of second messenger-dependent protein kinase A (PKA) and C (PKC) in the regulation of endogenous secretin receptor responsiveness in NG108-15 mouse neuroblastoma6rat glioma hybrid cells. 2 In whole cell cyclic AMP accumulation studies, activation of PKC either by phorbol 12-myristate 13-acetate (PMA) or by purinoceptor stimulation using uridine 5'-triphosphate (UTP) decreased secretin receptor responsiveness. PKC activation also inhibited forskolin-stimulated cyclic AMP accumulation but did not a ect cyclic AMP responses mediated by the prostanoid-IP receptor agonist iloprost, or the A 2 adenosine receptor agonist 5'-(N-ethylcarboxamido) adenosine (NECA). 3 In additivity experiments, saturating concentrations of secretin and iloprost were found to be additive in terms of cyclic AMP accumulation, whereas saturating concentrations of NECA and iloprost together were not. This suggests compartmentalization of G s -coupling components in NG108-15 cells and possible heterologous regulation of secretin receptor responsiveness at the level of adenylyl cyclase activation. 4 Cells exposed to the PKA inhibitor H-89, exhibited a time-dependent increase in secretin receptor responsiveness compared to control cells. This e ect was selective since cyclic AMP responses to forskolin, iloprost and NECA were not a ected by H-89 treatment. Furthermore, treatment with the protein synthesis inhibitor cycloheximide produced a time-dependent increase in secretin receptor responsiveness. 5 Together these results indicate that endogenous secretin receptor responsiveness is regulated by PKC, PKA and protein neosynthesis in NG108-15 cells.

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“…This process involves phosphorylation of the C‐terminal tail by PKC and thus seems to follow the general paradigm of GPCR signal attenuation. In contrast to this short‐term desensitization, cell systems naturally expressing IP‐R reveal a much slower time course of desensitization requiring 3–10 h [10–13]. The mechanism involved in this agonist‐induced long‐term attenuation is still poorly understood and was investigated in the present study.…”

Section: Introductionmentioning

confidence: 88%

Agonist‐induced long‐term desensitization of the human prostacyclin receptor

et al. 2000

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Phosphorylation of the human prostacyclin (PGI 2 ) receptor (hIP-R) by diacylglycerol-regulated protein kinase C (PKC) has been reported to be responsible for its rapid desensitization in HEK293 cells. In this study we demonstrate, that human fibroblasts reveal a much slower hIP-R desensitization kinetics, which was neither affected by stimulation nor inhibition of PKC by either phorbol 12-myristate-13-acetate or GF-109203X suggesting a different cellular mechanism. Although agonist-promoted sequestration of a C-terminally green fluorescent protein-tagged hIP-R was demonstrated, it did not account for the long-term desensitization. Concanavalin A did not abolish, but accelerated receptor desensitization kinetics. Resensitization of hIP-R involved receptor recycling and/or de novo synthesis of receptor protein, depending on the duration of prior desensitization. This is the first study investigating the mechanisms of hIP-R desensitization in intact human cells naturally expressing hIP-R. Our data suggest, that a hitherto unknown mechanism of hIP-R long-term desensitization, which is independent of receptor phosphorylation by conventional and novel type PKC isoforms or endocytosis, is a key event in regulating the cellular responsiveness to PGI 2 . ß

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The role of protein kinase A and protein kinase C in prostanoid IP receptor desensitization in NG108-15 cells

Cited by 9 publications

References 16 publications

Palmitoylation of the Human Prostacyclin Receptor

Palmitoylation of the Human Prostacyclin Receptor

Second messenger‐dependent protein kinases and protein synthesis regulate endogenous secretin receptor responsiveness

Agonist‐induced long‐term desensitization of the human prostacyclin receptor

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