Phorbol esters promote α1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Abstract

DDT1 MF-2 cells, which are derived from hamster vas deferens smooth muscle, contain α1-adrenergic receptors (54,800 ± 2700 sites per cell) that are coupled to stimulation of inositol phospholipid metabolism. Incubation of these cells with tumor-promoting phorbol esters, which stimulate calcium- and phospholipid-dependent protein kinase, leads to a marked attenuation of the ability of α1-receptor agonists such as norepinephrine to stimulate the turnover of inositol phospholipids. This turnover was measured by determining the 32P content of phosphatidylinositol and phosphatidic acid after prelabeling of the cellular ATP pool with 32P(i). These phorbol ester-treated cells also displayed a decrease in binding affinity of cellular α1 receptors for agonists with no change in antagonist affinity. By using affinity chromatography on the affinity resin Affi-Gel-A55414, the α1 receptors were purified ~300-fold from control and phorbol ester-treated 32P(i)-prelabeled cells. As assessed by NaDodSO4/polyacrylamide gel electrophoresis, the M(r) 80,000 α1-receptor ligand-binding subunit is a phosphopeptide containing 1.2 mol of phosphate per mol of α1 receptor. After phorbol ester treatment this increased to 3.6 mol of phosphate per mol of α1 receptor. The effect of phorbol esters on norepinephrine-stimulated inositol phospholipid turnover and α1-receptor phosphorylation showed the same rapid time course with a t( 1/2 )<min. These results indicate that calcium- and phospholipid-dependent protein kinase may play an important role in regulating the function of receptors that are coupled to the inositol phospholipid cycle by phosphorylating and deactivating them.

Detaljer

Författare
Externa organisationer
  • Duke University Medical Center
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Cell- och molekylärbiologi
Originalspråkengelska
Sidor (från-till)5651-5655
TidskriftProceedings of the National Academy of Sciences of the United States of America
Volym82
Utgivningsnummer17
StatusPublished - 1985 dec 1
PublikationskategoriForskning
Peer review utfördJa
Externt publiceradJa