Ca2+ release and Ca2+ influx in Chinese hamster ovary cells expressing the cloned mouse B2 bradykinin receptor: tyrosine kinase inhibitor-sensitive and- insensitive processes

Biochim Biophys Acta. 1997 Jan 10;1355(1):89-98. doi: 10.1016/s0167-4889(96)00126-7.

Abstract

A cDNA encoding a mouse B2 bradykinin (BK) receptor was stably transfected in Chinese hamster ovary (CHO) cells. In two resulting transformants, mouse B2 BK receptor was found to induce a twofold elevation in the inositol-1,4,5-trisphosphate level. In a pertussis toxin-insensitive manner, BK also produced a biphasic increase in the intracellular Ca2+ concentration ([Ca2+]i). The initial elevation in [Ca2+]i was abolished by thapsigargin pretreatment in Ca(2+)-free medium. The second phase was dependent on external Ca2+. The BK/inositol trisphosphate and thapsigargin-sensitive Ca2+ stores required extracellular Ca2+ for refilling. Ca2+ influx induced by BK and thapsigargin was confirmed by Mn2+ entry through Ca2+ influx pathways producing Mn2+ quenching. Genistein, a tyrosine kinase inhibitor, partially decreased the BK-induced [Ca2+]i increase during the sustained phase and the rate of Mn2+ entry. BK had essentially no effect on the intracellular cyclic AMP level. The results suggest that the mouse B2 BK receptor couples to phospholipase C in CHO cells and that its activation results in biphasic [Ca2+]i increases, by mobilization of intracellular Ca2+ and store-depletion-mediated Ca2+ influx, the latter of which is tyrosine phosphorylation dependent.

MeSH terms

  • Animals
  • Binding Sites
  • Bradykinin / metabolism
  • Bradykinin / pharmacology
  • CHO Cells
  • Calcium / metabolism*
  • Cloning, Molecular
  • Cricetinae
  • Cyclic AMP / metabolism
  • Enzyme Inhibitors / pharmacology
  • Genistein
  • Homeostasis
  • Inositol 1,4,5-Trisphosphate / metabolism
  • Isoflavones / pharmacology
  • Manganese / metabolism
  • Phosphorylation
  • Phosphotyrosine / metabolism
  • Protein-Tyrosine Kinases / antagonists & inhibitors
  • Protein-Tyrosine Kinases / metabolism*
  • Receptor, Bradykinin B2
  • Receptors, Bradykinin / genetics
  • Receptors, Bradykinin / metabolism*
  • Thapsigargin / pharmacology
  • Transfection

Substances

  • Enzyme Inhibitors
  • Isoflavones
  • Receptor, Bradykinin B2
  • Receptors, Bradykinin
  • Phosphotyrosine
  • Manganese
  • Thapsigargin
  • Inositol 1,4,5-Trisphosphate
  • Genistein
  • Cyclic AMP
  • Protein-Tyrosine Kinases
  • Bradykinin
  • Calcium