Expression and functional characterization of a pHis-tagged human bradykinin B2 receptor in COS-7 cells

Biol Chem. 2000 Apr;381(4):343-7. doi: 10.1515/BC.2000.045.

Abstract

A polyHis-tagged bradykinin (BK) B2 receptor (pHis-BKR) cDNA was constructed and expressed in COS-7 cells. The pHis-BKR is suitable for both immunoprecipitation and immunoblotting with anti-polyHis antibodies and can be easily purified using Ni-NTA columns. Immunochemical detection revealed a molecular mass of approximately 66 kDa. The pHis-BKR is capable of mediating BK-induced stimulation of inositol phosphate formation as well as of mitogen-activated protein kinase (MAPK) activity. Compared with the wild-type receptor (WT-BKR) the tagged receptor showed a slightly enhanced affinity towards BK but a reduced expression level. Despite these modified pharmacological properties the pHis-tagged BKR may be a useful tool for studying BKR modifications and signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenergic beta-Antagonists / metabolism
  • Affinity Labels / chemistry*
  • Animals
  • Anti-Inflammatory Agents, Non-Steroidal / metabolism
  • Binding Sites
  • Bradykinin / analogs & derivatives
  • Bradykinin / metabolism
  • Bradykinin Receptor Antagonists
  • COS Cells / metabolism*
  • Histidine / chemistry*
  • Histidine / immunology
  • Humans
  • Inhibitory Concentration 50
  • Kinetics
  • Peptides / chemistry
  • Phosphatidylinositols / metabolism
  • Protein Binding
  • Receptor, Bradykinin B2
  • Receptors, Bradykinin / genetics*
  • Receptors, Bradykinin / metabolism
  • Signal Transduction / drug effects
  • Transfection
  • Tritium

Substances

  • Adrenergic beta-Antagonists
  • Affinity Labels
  • Anti-Inflammatory Agents, Non-Steroidal
  • Bradykinin Receptor Antagonists
  • Peptides
  • Phosphatidylinositols
  • Receptor, Bradykinin B2
  • Receptors, Bradykinin
  • Tritium
  • Histidine
  • icatibant
  • Bradykinin