TY - JOUR
T1 - Evidence for α1-adrenergic receptor internalization in DDT1 MF-2 cells following exposure to agonists plus protein kinase C activators
AU - Cowlen, M. S.
AU - Toews, M. L.
N1 - Copyright:
Copyright 2004 Elsevier B.V., All rights reserved.
PY - 1988
Y1 - 1988
N2 - Agonist-induced sequestration and internalization of α1-adrenergic receptors were examined in DDT1 MF-2 cells. Pretreatment of cells with epinephrine or norepinephrine alone, but not with phorbol 12-myristate 13-acetate alone, resulted in a marked decrease in [3H]prazosin binding to intact cells at 4°. These pretreatments resulted in little or no change in the fraction of α1-adrenergic receptors exhibiting limited accessibility to the hydrophilic competing ligand epinephrine in short-time competition binding assays with intact cells and little or no change in the subcellular distribution of α1-adrenergic receptors between the plasma membrane and light vesicle compartments as assessed by sucrose density gradient centrifugation assays. Pretreatment with a combination of agonist plus phorbol 12-myristate 13-acetate resulted in a greater decrease in [3H]prazosin binding at 4° than was observed when cells were pretreated with agonist alone, induced the conversion of about half of cell surface α1-adrenergic receptors to a form exhibiting limited accessibility to epinephrine in short-term assays, and induced a shift of about half of α1-adrenergic receptors from the plasma membrane fraction to a light vesicle fraction on sucrose density gradients. A similar shift of α1-adrenergic receptors was observed on sucrose density gradients after exposure to agonist plus either mezerein or β-phorbol didecanoate, but not with agonist plus α-phorbol didecanoate, indicating involvement of protein kinase C. These results suggest that pretreatment with agonist alone induces the sequestration of α1-adrenergic receptors into a compartment that is inaccessible to [3H]prazosin at 4° but that is acccessible to hydrophilic ligands at 37° and remains associated with the plasma membrane. In contrast, α1-adrenergic receptors are apparently internalized from the plasma membrane to a separate compartment, presumably an intracellular vesicle, when cells are pretreated simultaneously with a combination of agonist plus a protein kinase C activator.
AB - Agonist-induced sequestration and internalization of α1-adrenergic receptors were examined in DDT1 MF-2 cells. Pretreatment of cells with epinephrine or norepinephrine alone, but not with phorbol 12-myristate 13-acetate alone, resulted in a marked decrease in [3H]prazosin binding to intact cells at 4°. These pretreatments resulted in little or no change in the fraction of α1-adrenergic receptors exhibiting limited accessibility to the hydrophilic competing ligand epinephrine in short-time competition binding assays with intact cells and little or no change in the subcellular distribution of α1-adrenergic receptors between the plasma membrane and light vesicle compartments as assessed by sucrose density gradient centrifugation assays. Pretreatment with a combination of agonist plus phorbol 12-myristate 13-acetate resulted in a greater decrease in [3H]prazosin binding at 4° than was observed when cells were pretreated with agonist alone, induced the conversion of about half of cell surface α1-adrenergic receptors to a form exhibiting limited accessibility to epinephrine in short-term assays, and induced a shift of about half of α1-adrenergic receptors from the plasma membrane fraction to a light vesicle fraction on sucrose density gradients. A similar shift of α1-adrenergic receptors was observed on sucrose density gradients after exposure to agonist plus either mezerein or β-phorbol didecanoate, but not with agonist plus α-phorbol didecanoate, indicating involvement of protein kinase C. These results suggest that pretreatment with agonist alone induces the sequestration of α1-adrenergic receptors into a compartment that is inaccessible to [3H]prazosin at 4° but that is acccessible to hydrophilic ligands at 37° and remains associated with the plasma membrane. In contrast, α1-adrenergic receptors are apparently internalized from the plasma membrane to a separate compartment, presumably an intracellular vesicle, when cells are pretreated simultaneously with a combination of agonist plus a protein kinase C activator.
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M3 - Article
C2 - 2901664
AN - SCOPUS:0023813416
SN - 0026-895X
VL - 34
SP - 340
EP - 346
JO - Molecular Pharmacology
JF - Molecular Pharmacology
IS - 3
ER -